Pharmaceutical compositions and related methods of delivery

ABSTRACT

The pharmaceutical compositions described herein include a suspension which comprises an admixture in solid form of a therapeutically effective amount of a therapeutic agent and at least one salt of a medium chain fatty acid and a hydrophobic medium, e.g. castor oil or glyceryl tricaprylate or a mixture thereof. The pharmaceutical compositions described herein contain medium chain fatty acid salts and are substantially free of alcohols. The pharmaceutical compositions may be encapsulated in a capsule. Methods of treating or preventing diseases by administering such compositions to affected subjects are also disclosed.

CLAIM OF PRIORITY

The present application is a continuation of U.S. application Ser. No.14/188,139, filed Feb. 24, 2014, which is a continuation of U.S.application Ser. No. 12/981,036, filed Dec. 29, 2010, which is acontinuation of U.S. application Ser. No. 12/561,738, filed Sep. 17,2009 which claims priority from U.S. Ser. No. 61/097,716, filed Sep. 17,2008, U.S. Ser. No. 61/141,686, filed Dec. 31, 2008, and U.S. Ser. No.61/161,387, filed Mar. 18, 2009, each of which is incorporated byreference in its entirety.

FIELD OF THE TECHNOLOGY

The present invention relates generally to pharmaceutical compositionsenabling improved delivery e.g. oral delivery and methods of using suchcompositions.

BACKGROUND

Techniques enabling efficient transfer of a substance of interest acrossa biological barrier are of considerable interest in the fields ofbiotechnology and medicine. For example, such techniques may be used forthe transport of a variety of different substances across a biologicalbarrier regulated by tight junctions (i.e., the mucosal epithelia, whichinclude the intestinal and respiratory epithelia, and the vascularendothelia, which include the blood-brain barrier, nasal membrane,cornea and other eye membranes, and genito-urinary membranes). Inparticular there is great interest in oral delivery of therapeuticagents to avoid the use of more invasive means of administration andhence improve patient convenience and compliance.

Diverse drug delivery vehicles have been employed, among them liposomes,lipidic or polymeric nanoparticles, and microemulsions. These haveimproved the oral bioavailability of certain drugs, mostly by theprotective effect they offer. However, for most relevant drugs,bioavailability remains very low and fails to achieve the minimaltherapeutic goals.

Hence, a need exists for an efficient, specific, non-invasive, low-riskmeans to target various biological barriers for the non invasivedelivery of various therapeutic agents such as peptides andpolypeptides, macromolecule drugs and other therapeutic agents whichinclude small molecules with low bioavailability.

SUMMARY

The inventors of the present invention have discovered that theabsorption of certain therapeutic agents in a subject can be improvedwhen administered in a composition described herein. For example, atherapeutic agent administered in a formulation in accordance with oneor more embodiments exhibits an improved bioavailability (BA) relativeto the same therapeutic agent administered via a similar route but in acomposition substantially free of the medium chain fatty acid saltcomponent described herein or having a lower amount of the medium chainfatty acid salt component described herein. Such improvement in relativeBA may be on the order of at least about 1.5-, 2-, 3-, 5-, 10-, 50- or100-fold. In some aspects, a composition described herein improves theabsorption in the gastrointestinal (GI) tract of a therapeutic agentthat is generally characterized by low or zero oral bioavailabilityand/or absorption. These therapeutic agents may have low or zerobioavailability, e.g., in aqueous solution, and in other oralformulations known in the art. In at least one aspect, a compositiondescribed herein improves bioavailability by enhancing the GIwall/barrier permeability to the drug molecules. For example, acomposition described herein may facilitate absorption by permeating theGI wall/barrier primarily via unsealing of the tight junctions betweenGI epithelial cells, although it may also work by transcellularabsorption.

The present inventors have devised a process for producing apharmaceutical composition (bulk drug product) which involves preparinga water soluble composition comprising a therapeutically effectiveamount of at least one therapeutic agent and a medium chain fatty acidsalt (and other ingredients—see below), drying (e.g. by lyophilization)the water soluble composition to obtain a solid powder, and suspendingthe lyophilized material (the solid powder) in a hydrophobic (oily)medium, preferably castor oil or glyceryl tricaprylate (including otheringredients e.g. PVP and surfactants and viscosity modifiers—see below),to produce a suspension containing in solid form the therapeutic agentand the medium chain fatty acid salt, thereby producing the bulk drugproduct, which must contain at least 10% by weight of medium chain fattyacid salt. The solid form may comprise a particle (e.g., consistsessentially of particles, or consists of particles. The particle may beproduced by lyophilization or by granulation. The bulk drug product maythen be encapsulated in capsules which will be coated by a pH sensitivecoating and may be used for oral delivery. A typical process forproducing the claimed formulation is shown in FIG. 1, where insulin isexemplified as the active pharmaceutical ingredient (API) and the mediumchain fatty acid salt is sodium octanoate (Na—C8), also termed sodiumcaprylate.

The present invention demonstrates delivery of the product to theintestine, which is a model for oral delivery, and from there to thebloodstream with high bioavailability.

Thus in one aspect the invention features a composition. The compositionincludes a therapeutic agent and a medium chain fatty acid saltassociated with a substantially hydrophobic medium, preferably castoroil, wherein the therapeutic agent and the medium chain fatty acid saltthereof are in solid form, e.g. in the same solid form such as aparticle, obtained by drying from an aqueous medium, e.g. bylyophilizing the aqueous medium, and wherein the medium chain fatty acidsalt is present at 10% by weight or more, preferably 12-15%, e.g., about12%, about 13%, about 14%, or about 15% or about 16%, or about 17%, andwherein the composition contains other ingredients (as described herein)but is substantially free of a “membrane fluidizing agent”. “Membranefluidizing agents” are defined as various linear, branched, aromatic andcyclic medium chain alcohols, in particular geraniol and octanol.

The present compositions of the invention are not emulsions. Almost allof the present compositions are oily suspensions and the amount of waterin the compositions is very low; a few of the present compositions whichare not suspensions incorporate a high amount (about 78% octanoic acid)and are solutions.

In the compositions of the invention, the therapeutic agent and mediumchain fatty acid salt are in intimate contact with the substantiallyhydrophobic medium. For example, a powder comprising the therapeuticagent and medium chain fatty acid salt is coated, immersed or suspendedin the substantially hydrophobic medium.

During the production process the aqueous medium which contains thetherapeutic agent and the medium chain fatty acid salt and the otheringredients is dried (e.g. by lyophilization) to obtain the hydrophilicfraction which is a powder (e.g., a solid form comprising a plurality ofparticles), and a particle in that powder contains all the ingredientsi.e. the therapeutic agent and medium chain fatty acid salt are togetherin a single particle. The solid form may be, for example, a granulatedparticle or a lyophilized particle.

In some embodiments, the therapeutic agent is selected from the groupconsisting of peptides, polysaccharides, polynucleotides, and smallmolecules. The therapeutic agent may be a protein. For example, thetherapeutic agent may be insulin. In other embodiments, the therapeuticagent is a polynucleotide e.g. DNA or RNA compound. In some embodiments,the therapeutic agent is a small molecule, a poorly soluble drug, or ahighly crystalline drug. The therapeutic agent may be a growth hormone.In at least one embodiment, the therapeutic agent is teriparatide. Insome embodiments, the therapeutic agent may be leuprolide or alendronateor octreotide.

In some embodiments, the composition includes a plurality of mediumchain fatty acid salts and derivatives thereof. For example, the solidparticle may further include a plurality of medium chain fatty acidsalts and derivatives thereof.

In some embodiments, the medium chain fatty acid salt is selected fromthe group consisting of sodium hexanoate, sodium heptanoate, sodiumoctanoate, sodium nonanoate, sodium decanoate, sodium undecanoate,sodium dodecanoate, sodium tridecanoate, and sodium tetradecanoate or acombination thereof. In accordance with one or more embodiments, thecomposition is substantially free of sodium dodecanoate, sodiumtridecanoate, and sodium tetradecanoate. In some embodiments, the mediumchain fatty acid is sodium octanoate and the sodium octanoate is presentat a concentration of above 10% e.g. about 11% to about 50%weight/weight (wt/wt).

In some embodiments, the substantially hydrophobic medium comprises atriglyceride. For example, the triglyceride may be selected from thegroup consisting of glyceryl tributyrate, glyceryl monooleate, glycerylmonocaprylate and glyceryl tricaprylate.

In some embodiments, the substantially hydrophobic medium comprisesmineral oil, castor oil, olive oil, corn oil, coconut oil, peanut oil,soybean oil, cotton seed oil, sesame oil or canola oil, or combinationsthereof.

In some embodiments the water-soluble composition contains a mediumchain fatty acid salt and the hydrophobic medium contains thecorresponding medium chain fatty acid; in some particular embodimentsthe medium chain fatty acid salt is a salt of octanoic acid such assodium octanoate and the medium chain fatty acid is octanoic acid.

In some embodiments the water-soluble composition contains a mediumchain fatty acid salt and the hydrophobic medium contains thecorresponding medium chain monoglyceride or the corresponding mediumchain triglyceride or a combination thereof; in some particularembodiments the medium chain fatty acid salt is sodium octanoate and themonoglyceride is glyceryl monocaprylate and the triglyceride is glyceryltricaprylate.

In some embodiments, the composition further includes one or moreexcipients. The excipients may be a salt e.g MgCl₂ or an aminecontaining compound or mannitol. In some embodiments, the excipient isin the same solid form as the therapeutic agent.

In some embodiments the excipient is a stabilizer. The inventorsunexpectedly found that although polyvinylpyrolidine (PVP) in particularPVP-12 is known in the art as a stabilizer, in formulations of theinvention it serves to increase the effect of the permeability enhanceron absorbance of the therapeutic agent.

In some embodiments, the composition further includes one or moresurfactants. For example, the surfactant may be selected from the groupconsisting of sorbitan monopalmitate (Span-40®), polyoxyethylenesorbitanmonooleate (Tween80), lecithin, and glyceryl monooleate (GMO). In one ormore embodiments, the surfactant comprises from about 0.1% to about 6%by weight of the composition.

In preferred embodiments, the composition is an oral dosage form. Forexample, the composition may be filled in a hard or soft capsule. Insome embodiments, the composition is in the form of a suppository. Inaccordance with one or more embodiments, the composition may be in theform of an enema fleet.

In some embodiments, the bioavailability of the therapeutic agent, whenadministered to a subject, is at least 1.5-2% relative to parenteral(subcutaneous or intravenous) administration. In some embodiments, thecomposition, when administered to a subject, provides above 2%, above3%, above 5%, above 10%, or above 20% or above 30% absorption of thetherapeutic agent across a biological barrier. The levels of absorptionachieved produce the therapeutic levels needed for the indicationconcerned.

In one aspect, the invention features a method of treating a disorder ina subject. The method includes administering to the subject any one ofthe compositions described herein.

In some embodiments, the composition is administered orally. In otherembodiments, the composition is administered rectally, sublingually orvia buccal administration.

In some embodiments, the disorder may be anemia. In accordance with oneor more embodiments, the disorder is osteoporosis. The disorder may befemale infertility. In other embodiments, the disorder is growth failureor growth hormone deficiency. In at least one embodiment, the disorderis HIV-related weight loss or wasting, acromegaly or diabetes.

In some embodiments the therapeutic agent is octreotide and the disorderis acromegaly, abnormal GI motility, gastroparesis, diarrhea or portalhypertension.

In some embodiments, the method may include encapsulating the suspensionto form a capsule. The method may further include coating the capsule.

In some embodiments, the method may include providing instructions toadminister the capsule to a subject. The instructions may relate toadministering the capsule to a subject for any indication describedherein. In one aspect, the invention features capsules provided withinstructions relating to administering the capsule to a subject for anyindication described herein.

Still other aspects, embodiments, and advantages of these exemplaryaspects and embodiments, are discussed in detail below. Moreover, it isto be understood that both the foregoing information and the followingdetailed description are merely illustrative examples of various aspectsand embodiments, and are intended to provide an overview or frameworkfor understanding the nature and character of the claimed aspects andembodiments. The accompanying drawings are included to provideillustration and a further understanding of the various aspects andembodiments, and are incorporated in and constitute a part of thisspecification. The drawings, together with the remainder of thespecification, serve to explain principles and operations of thedescribed and claimed aspects and embodiments.

Throughout this application, various publications, including UnitedStates patents, are referenced by author and year and patents andapplications by number. The disclosures of these publications andpatents and patent applications in their entireties are herebyincorporated by reference into this application in order to more fullydescribe the state of the art to which this invention pertains.

BRIEF DESCRIPTION OF THE DRAWINGS

Various aspects of at least one embodiment are discussed below withreference to the accompanying Figures. In the Figures, which are notintended to be drawn to scale, each identical or nearly identicalcomponent that is illustrated in various figures is represented by alike numeral. For purposes of clarity, not every component may belabeled in every drawing. The Figures are provided for the purposes ofillustration and explanation and are not intended as a definition of thelimits of the invention. In the Figures:

FIG. 1 presents a process for production of an insulin formulation of acomposition in accordance with one or more embodiments as referenced inthe accompanying Examples;

FIGS. 2-5 present data referenced in accompanying Examples 3 through 6;

FIG. 6 presents data referenced in accompanying Example 8;

FIG. 7 presents molecular weight marker permeability data referenced inaccompanying Example 33;

FIG. 8 presents time-course permeability data referenced in accompanyingExample 34; and

FIGS. 9 and 10 present data relating to administration of octreotide tomonkeys referenced in accompanying Example 35.

DETAILED DESCRIPTION

The compositions described herein can be administered to a subject toprovide for improved bioavailability of a therapeutic agent.

Pharmaceutical Compositions:

The pharmaceutical compositions described herein include a therapeuticagent and a medium chain fatty acid salt in intimate contact orassociation with a substantially hydrophobic medium. For example, thetherapeutic agent and the medium chain fatty acid or derivative thereofmay be coated, suspended, sprayed by or immersed in a substantiallyhydrophobic medium forming a suspension. The compositions of theinvention are not emulsions. Almost all of the compositions are oilysuspensions and the amount of water in the compositions is very low; afew of the present compositions which are not suspensions incorporate ahigh amount (about 78% octanoic acid) and are solutions by visualanalysis. The suspension may be a liquid suspension incorporating solidmaterial, or a semi-solid suspension incorporating solid material (anointment).

Many of the compositions described herein comprise a suspension whichcomprises an admixture of a hydrophobic medium and a solid form whereinthe solid form comprises a therapeutically effective amount of atherapeutic agent and at least one salt of a medium chain fatty acid,and wherein the medium chain fatty acid salt is present in thecomposition at an amount of 10% or more by weight. The solid form maycomprise a particle (e.g., consist essentially of particles, or consistof particles). The particle may be produced by lyophilization or bygranulation. In some embodiments, preferably after milling, 90% (v/v) ofthe particles are below 130 microns, and 50% (v/v) of the particles arebelow 45 microns.

A cargo compound is a therapeutic agent (e.g. insulin) or a testcompound (e.g. high molecular weight dextran) which is formulated asdescribed herein within the compositions of the invention.

The inventors were particular to include in many of the compositions ofthe invention only excipients which are generally recognized as safe,based on available data on human use, animal safety and regulatoryguidelines (e.g. GRAS excipients). Some compositions of the inventionmay have other types of excipients (e.g. non-GRAS). In some embodimentsthe compositions of the invention have amounts of excipients that arewithin the maximum daily doses as noted in such available data for eachspecific excipient.

The medium chain fatty acid salt may generally facilitate or enhancepermeability and/or absorption of the therapeutic agent. In someembodiments the medium chain fatty acid salts include derivatives ofmedium chain fatty acid salts. The therapeutic agent and the mediumchain fatty acid salt are in solid form, for example, a solid particlesuch as a lyophilized particle, granulated particle, pellet ormicro-sphere. In preferred embodiments, the therapeutic agent and themedium chain fatty acid salt are both in the same solid form, e.g., bothin the same particle. In other embodiments, the therapeutic agent andthe medium chain fatty acid salt may each be in a different solid form,e.g. each in a distinct particle. The compositions described herein aresubstantially free of any “membrane fluidizing agents” defined aslinear, branched, aromatic and cyclic medium chain alcohols, inparticular geraniol and octanol. For example the compositions preferablyinclude no membrane fluidizing agents but certain embodiments mayinclude for example less than 1% or less than 0.5% or less than 0.1% byweight of membrane fluidizing agents.

Unlike emulsions, where water is an essential constituent of theformulation, the compositions described herein provide a solid form suchas a particle containing the therapeutic agent, which is then associatedwith the hydrophobic (oily) medium. The amount of water in thecompositions is generally less than 3% by weight, usually less thanabout 2% or about 1% or less by weight.

The compositions described herein are suspensions which comprise anadmixture of a hydrophobic medium and a solid form wherein the solidform comprises a therapeutically effective amount of a therapeutic agentand at least one salt of a medium chain fatty acid. The solid form maybe a particle (e.g., consist essentially of particles, or consist ofparticles). The particle may be produced by lyophilization or bygranulation. The medium chain fatty acid salt is generally present inthe compositions described herein at an amount of 10% or more by weight.In certain embodiments the medium chain fatty acid salt is present inthe composition at an amount of 10%-50%, preferably 11%-18% or about11%-17% or 12%-16% or 12%-15% or 13%-16% or 13%-15% or 14%-16% or14%-15% or 15%-16% or most preferably 15% or 16% by weight, and themedium chain fatty acid has a chain length from about 6 to about 14carbon atoms preferably 8, 9 or 10 carbon atoms.

In some embodiments in the compositions described above, the solid formincluding the therapeutic agent also includes a stabilizer (e.g., astabilizer of protein structure). Stabilizers of protein structure arecompounds that stabilize protein structure under aqueous or non-aqueousconditions or can reduce or prevent aggregation of the therapeuticagent, for example during a drying process such as lyophilization orother processing step. Stabilizers of structure can be polyanionicmolecules, such as phytic acid, polyvalent ions such as Ca, Zn or Mg,saccharides such as a disaccharide (e.g., trehalose, maltose) or anoligo or polysaccharide such as dextrin or dextran, or a sugar alcoholsuch as mannitol, or an amino acid such as glycine, or polycationicmolecules, such as spermine, or surfactants such as polyoxyethylenesorbitan monooleate (Tween 80) or pluronic acid. Uncharged polymers,such as mannitol, methyl cellulose and polyvinyl alcohol, are alsosuitable stabilizers.

Although polyvinylpyrrolidone (PVP) is known in the art as a stabilizer,the inventors unexpectedly found that, in the compositions of theinvention described herein, PVP, in particular PVP-12, serves toincrease the effect of the permeability enhancer in a synergisticmanner; furthermore, increasing the level of PVP-12 to 10% increased theabsorption of the therapeutic agent into the blood due to the improvedactivity of the formulations. The inventors demonstrated that dextranhad a similar (but lower) effect as PVP did. Other matrix formingpolymers have a similar effect.

In some embodiments, such as when the therapeutic agent is a smallmolecule, a bulking agent may be added, for example, mannitol or glycin.

In certain embodiments of the compositions described herein thetherapeutic agent is a protein, a polypeptide, a peptide, aglycosaminoglycan, a small molecule, a polysaccharide or apolynucleotide inter alia, such as octreotide, growth hormone,parathyroid hormone, parathyroid hormone amino acids 1-34 [PTH(1-34)termed teriparatide], a low molecular weight heparin or fondaparinuxinter alia. Low molecular weight heparins are defined as heparin saltshaving an average molecular weight of less than 8000 Da and for which atleast 60% of all chains have a molecular weight less than 8000 Da.

In a particular embodiment of the compositions described herein the saltof the fatty acid is sodium octanoate and the hydrophobic medium iscastor oil; in another particular embodiment the composition furthercomprises glyceryl monooleate and sorbitan monopalmitate or glycerylmonocaprylate and glyceryl tricaprylate and polyoxyethylenesorbitanmonooleate; in another particular embodiment the composition furthercomprises glyceryl tributyrate, lecithin, ethylisovalerate and at leastone stabilizer. In particular embodiments the therapeutic agent isoctreotide, growth hormone, parathyroid hormone, teriparatide,interferon-alfa (IFN-α), a low molecular weight heparin, fondaparinux,siRNA, somatostatin and analogs (agonists) thereof includingpeptidomimetics, exenatide, vancomycin or gentamicin inter alia.

Therapeutic Agents:

The pharmaceutical compositions described herein can be used with avariety of therapeutic agents (also termed active pharmaceuticalingredient=API). In some embodiments, the pharmaceutical compositionincludes a plurality of therapeutic agents (effectors). The therapeuticagents can either be in the same solid form (e.g., in the sameparticle), or the therapeutic agents can each be in an independent solidform (e.g., each in different particles. In some embodiments, thetherapeutic agent is in the form of a particle, for example, agranulated or solid particle. The particle is associated with or is inintimate contact with a substantially hydrophobic medium, for example, ahydrophobic medium described herein.

Therapeutic agents that can be used in the compositions described hereininclude any molecule or compound serving as, for example, a biological,therapeutic, pharmaceutical, or diagnostic agent including an imagingagent. The therapeutic agents include drugs and other agents including,but not limited to, those listed in the United States Pharmacopeia andin other known pharmacopeias. Therapeutic agents are incorporated intothe formulations of the invention without any chemical modification.Therapeutic agents include proteins, polypeptides, peptides,polynucleotides, polysaccharides and small molecules.

The term “small molecule” is understood to refer to a low molecularweight organic compound which may be synthetically produced or obtainedfrom natural sources and typically has a molecular weight of less than2000 Da, or less than 1000 Da or even less than 600 Da e.g. less than orabout 550 Da or less than or about 500 Da or less than or about 400 Da;or about 400 Da to about 2000 Da; or about 400 Da to about 1700 Da.Examples of small molecules are ergotamine (molecular weight=582 Da),fondaparinux (molecular weight=1727 Da), leuprolide (molecularweight=1209 Da), vancomycin (molecular weight=1449 Da), gentamicin(molecular weight=478 Da) and doxorubicin (molecular weight=544).

The term “polynucleotide” refers to any molecule composed of DNAnucleotides, RNA nucleotides or a combination of both types whichcomprises two or more of the bases guanidine, citosine, timidine,adenine, uracil or inosine, inter alia. A polynucleotide may includenatural nucleotides, chemically modified nucleotides and syntheticnucleotides, or chemical analogs thereof and may be single-stranded ordouble-stranded. The term includes “oligonucleotides” and encompasses“nucleic acids”.

By “small interfering RNA” (siRNA) is meant an RNA molecule(ribonucleotide) which decreases or silences (prevents) the expressionof a gene/mRNA of its endogenous or cellular counterpart. The term isunderstood to encompass “RNA interference” (RNAi), and “double-strandedRNA” (dsRNA).

By “polypeptide” is meant a molecule composed of covalently linked aminoacids and the term includes peptides, polypeptides, proteins andpeptidomimetics. A peptidomimetic is a compound containing non-peptidicstructural elements that is capable of mimicking the biologicalaction(s) of a natural parent peptide. Some of the classical peptidecharacteristics such as enzymatically scissile peptidic bonds arenormally not present in a peptidomimetic.

The term “amino acid” refers to a molecule which consists of any one ofthe 20 naturally occurring amino acids, amino acids which have beenchemically modified or synthetic amino acids.

By “polysaccharide” is meant a linear or branched polymer composed ofcovalently linked monosaccharides; glucose is the most commonmonosaccharide and there are normally at least eight monosaccharideunits in a polysaccharide and usually many more. Polysaccharides have ageneral formula of Cx(H2O)y where x is usually a large number between200 and 2500. Considering that the repeating units in the polymerbackbone are often six-carbon monosaccharides, the general formula canalso be represented as (C6H10O5)n where 40≦n≦3000 i.e. there arenormally between 40 and 3000 monosaccharide units in a polysaccharide.

A “glycosaminoglycan” is a polysaccharide that contains amino containingsugars.

Exemplary anionic therapeutic agents include polynucleotides fromvarious origins, and particularly from human, viral, animal, eukaryoticor prokaryotic, plant, or synthetic origin, etc including systems fortherapeutic gene delivery. A polynucleotide of interest may be of avariety of sizes, ranging from, for example, a simple trace nucleotideto a gene fragment, or an entire gene. It may be a viral gene or aplasmid. Exemplary polynucleotides serving as therapeutic agents includespecific DNA sequences (e.g., coding genes), specific RNA sequences(e.g., RNA aptamers, antisense RNA, short interfering RNA (siRNA) or aspecific inhibitory RNA (RNAi)), poly CPG, or poly I:C syntheticpolymers of polynucleotides.

Alternatively, the therapeutic agent can be a protein, such as, forexample, an enzyme, a hormone, an incretin, a proteoglycan, a ribozyme,a cytokine, a peptide, an apolipoprotein, a growth factor, a bioactivemolecule, an antigen, or an antibody or fragment(s) thereof, etc. Thepeptide can be a small peptide e.g. from about 2 to about 40 aminoacids, examples include fibrinogen-receptor antagonists (RGD-containingpeptides which are tetrapeptides having an average molecular weight ofabout 600. Exemplary peptides are somatostatin and analogs thereof e.g.octreotide and lanreotide (Somatuline) which are both cyclicoctapeptides and pasireotide (SOM-230) which is a cyclic hexapeptide(Weckbecker et al, 2002, Endocrinology 143(10) 4123-4130; Schmid, 2007,Molecular and Cellular Endocrinology 286, 69-74). Other exemplarypeptides are glatiramer acetate (Copaxone®) which is a tetrapeptide,terlipressin which is a 12 amino acid peptide analog (agonist) of lysinevasopressin (ADH) and exenatide, a 39 amino acid peptide which is anincretin mimetic agent, and other analogs of glucagon-likepeptide-1(GLP-1). (Byetta® is the trade name for exenatide (Eli Lillyand Company/Amylin Pharmaceuticals, Inc.). Other peptides includedalargin which is a hexapeptide, and kyotorphin which is a dipeptide.Peptides include growth hormone releasing peptides which are peptides ofabout 12 amino acids or less; see for example peptides disclosed in U.S.Pat. No. 4,411,890 (Momany) and U.S. Pat. No. 4,839,344 (Bowers et al)

Examples of other peptides which can be used in the practice of thisinvention are those disclosed in U.S. Pat. No. 4,589,881 (30 or moreamino acid residues) of Pierschbacher et al; U.S. Pat. No. 4,544,500(20-30 residues) of Bittle et al; and EP0204480 (>34 residues) ofDimarchi et al and teriparatide. In some embodiments, the therapeuticagent can include a polysaccharide, such as a glycosaminoglycan.Exemplary glycosaminoglycans include heparin, heparin derivatives,heparan sulfate, chondroitin sulfate, dermatan sulfate, and hyaluronicacid. Examples of heparin derivatives include, but are not limited to,low molecular weight heparins such as enoxaparin, dalteparin andtinzaparin. A therapeutic agent with a heparin-like effect isfondaparinux.

Other examples of therapeutic agents include, but are not limited tohormones such as insulin, erythropoietin (EPO), glucagon-like peptide 1(GLP-1), melanocyte stimulating hormone (alfa-MSH), parathyroid hormone(PTH), teriparatide, growth hormone (GH), leuprolide, leuprolideacetate, factor VIII, growth hormone releasing hormone (GHRH), peptideYY amino acids 3-36 (PYY₍₃₋₃₆₎), calcitonin, somatotropin, somatostatin,somatomedin, interleukins such as interleukin-2 (IL-2),alfa-1-antirypsin, granulocyte/monocyte colony stimulating factor(GM-CSF), granulocyte colony stimulating factor (G-CSF), T20,testosterone, interferons such as interferon-alfa (IFN-α) IFN-β andIFN-γ, luteinizing hormone (LH), follicle-stimulating hormone (FSH),human chorionic gonadotropin (hCG), enkephalin, dalargin, kyotorphin,basic fibroblast growth factor (bFGF), hirudin, hirulog, luteinizinghormone releasing hormone (LHRH), gonadotropin releasing hormone (GnRH)analog, brain-derived natriuretic peptide (BNP), tissue plasminogenactivator (TPA), oxytocin, and analogs and combinations thereof.

Other examples of therapeutic agents include, but are not limited toanalgesic agents, anti-migraine agents, anti-coagulant agents,anti-emetic agents, cardiovascular, anti-hypertensive and vasodilatoragents, sedatives, narcotic antagonists, chelating agents, anti-diureticagents and anti-neoplastic agents.

Analgesics include, but are not limited to, fentanyl, sufentanil,butorphanol, buprenorphine, levorphanol, morphine, hydromorphone,hydrocodeine, oxymorphone, methadone, lidocaine, bupivacaine,diclofenac, naproxen, paverin, and analogs thereof. Anti-migraine agentsinclude, but are not limited to naratriptan, naproxen, almotriptan,butalbital, frovatriptan, sumatriptan, rizatriptan, acetaminophen,isometheptene, butorphanol, dichloralphenazone, ergot alkaloids such asdihydroergotamine and ergotamine, nonsteroidal anti-inflammatory drugs(NSAIDs) such as ketoprofen and ketorolac, eletriptan, butorphanol,topiramate, zolmitriptan, caffeine, aspirin and codeine, and analogs andcombinations thereof.

Anti-coagulant agents include, but are not limited to heparin, hirudin,low molecular weight heparins and analogs thereof and fondaparinux.Anti-emetic agents include but are not limited to scopolamine,ondansetron, domperidone, etoclopramide, and analogs thereof.Cardiovascular, anti-hypertensive and vasodilator agents include, butare not limited to, diltiazem, clonidine, nifedipine, verapamil,isosorbide-5-mononitrate, organic nitrates, nitroglycerine and analogsthereof. Sedatives include, but are not limited to, benzodiazeines,phenothiozines and analogs thereof. Narcotic antagonists include, butare not limited to, naltrexone, naloxone and analogs thereof. Chelatingagents include, but are not limited to deferoxamine and analogs thereof.Anti-diuretic agents include, but are not limited to, desmopressin,vasopressin and analogs (agonists) thereof such as terlipressin; thetrade name of terlipressin is Glypressin®. Anti-neoplastic agentsinclude, but are not limited to, 5-fluorouracil, bleomycin, vincristine,procarbazine, temezolamide, 6-thioguanine, hydroxyurea, cytarabine,cyclophosphamide, doxorubicin, vinca alkaloid, epirubicin, etoposide,ifosfamide, carboplatin and other platinum based antineoplastic drugs(such as carboplatin (Paraplatin®, tetraplatin, oxaliplatin, aroplatinand transplatin), vinblastine, vinorelbine, chlorambucil, busulfan,mechlorethamine, mitomycin, dacarbazine, thiotepa, daunorubicin,idarubicin, mitoxantrone, esperamicin A1, dactinomycin, plicamycin,carmustine, lomustine (CCNU), tauromustine, streptozocin, melphalan,dactinomycin, procarbazine, dexamethasone, prednisone,2-chlorodeoxyadenosine, cytarabine, docetaxel, fludarabine, gemcitabine,herceptin, hydroxyurea, irinotecan, methotrexate, rituxin, semustine,tomudex and topotecan, taxol and taxol-like compounds and analogs andcombinations thereof.

Additional examples of therapeutic agents include, but are not limitedto coagulation factors and neurotrophic factors, anti-TNF antibodies andfragments of TNF receptors.

Therapeutic agents also include pharmaceutically active agents selectedfrom the group consisting of vitamin B12, a bisphosphonate (e.g.,disodium pamidronate, alendronate, etidronate, tiludronate, risedronate,zoledronic acid, sodium clodronate, or ibandronic acid), taxol,caspofungin, or an aminoglycoside antibiotic. Additional therapeuticagents include a toxin, or an antipathogenic agent, such as anantibiotic (e.g. vancomycin), an antiviral, an antifungal, or ananti-parasitic agent. The therapeutic agent can itself be directlyactive or can be activated in situ by the composition, by a distinctsubstance, or by environmental conditions.

In some embodiments, the composition can include a plurality oftherapeutic agents (combination drugs). For example, the composition caninclude Factor VIII and vWF, GLP-1 and PYY, IFN-α and nucleotideanalogues (i.e. ribavirin), and alendronate or insulin and GLP-1.

In some embodiments, the composition can include a small molecule and apeptide or protein. Exemplary combinations include a combination ofIFN-α and nucleotide analogues (i.e. ribavirin) for the treatment ofhepatitis C, teriparatide and alendronate for treatment of bonedisorders, a combination of GH plus the medications for HIV therapy(e.g., HAART) to simultaneously treat the viral infection and theaccompanying HIV lipodystrophy or AIDS wasting side effects.Combinations of two small molecules can be used when one of themgenerally has poor absorption or bioavailability even if the othergenerally has effective absorption or bioavailability, such as someantibiotics (e.g., a combination of vancomycin and an aminoglycosidesuch as gentamicin. Exemplary combinations for the treatment andprevention of metabolic disorders such as diabetes and obesity alsoinclude combination of insulin and metformin, insulin and rosiglitazone,GLP-1 (or exenatide) and metformin, and GLP-1 (or exenatide) androsiglitazone.

Indications and conditions which may be treated by fondaparinuxformulated as described herein include deep vein thrombosis, hip or kneereplacement, and bed-bound patients.

In some embodiments of the compositions described herein, thecomposition includes a combination of a protein or peptide with smallmolecules that either do or do not have good absorption orbioavailability. For example, a composition can include at least onetherapeutic agent that may generally be characterized as poorlyabsorbable or poorly bioavailable. The composition can also be used forthe administration of therapeutic agents that are absorbed in thestomach and/or intestine, but cause irritation to the stomach and/orintestine and therefore are difficult to tolerate. In such a situation,a subject could benefit if the bioavailability of the therapeutic agentwere enhanced or if more of the therapeutic agent were absorbed directlyinto the blood stream; if less therapeutic agent is administered therewill clearly be less chance of causing irritation to the stomach and/orintestine. Thus compositions of the invention are envisaged whichcomprises therein two or more therapeutic agents.

In general, the composition may include from about 0.01% to about 50% byweight of the therapeutic agent e.g. about 0.01, 0.02 0.05, 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, or 50% by weight. Themaximum included in the composition is often in the range of about6%-33% by weight of the therapeutic agent.

In some embodiments of the compositions described herein, the solid formincluding the therapeutic agent also includes a stabilizer (e.g., astabilizer of protein structure). Stabilizers of protein structure arecompounds that stabilize protein structure under aqueous or non-aqueousconditions or can reduce or prevent aggregation of the therapeuticagent, for example during a drying process such as lyophilization orother processing step. Stabilizers of structure can be polyanionicmolecules, such as phytic acid, polyvalent ions such as Ca, Zn or Mg,saccharides such as a disaccharide (e.g., trehalose, maltose) or anoligo or polysaccharide such as dextrin or dextran, or a sugar alcoholsuch as mannitol, or an amino acid such as glycine, or polycationicmolecules, such as spermine, or surfactants such as Tween 80 or Span 40or pluronic acid. Uncharged polymers, such as methyl cellulose andpolyvinyl alcohol, are also suitable stabilizers.

Medium Chain Fatty Acid Salt:

The compositions described herein include the salt of a medium chainfatty acid or a derivative thereof in a solid form. For example, thesalt of the medium chain fatty acid is in the form of a particle such asa solid particle. In some embodiments, the particle may be characterizedas a granulated particle. In at least some embodiments, the solid formmay generally result from a spray drying or evaporation process. Inpreferred embodiments, the salt of the medium chain fatty acid is in thesame particle as the therapeutic agent. For example, the therapeuticagent and the salt of the medium chain fatty acid can be preparedtogether by first preparing a solution such as an aqueous solutioncomprising both the therapeutic agent and the salt of the medium chainfatty acid and co-lyophilizing the solution to provide a solid form orparticle that comprises both the therapeutic agent and the salt of themedium chain fatty acid (and other ingredients). As described above, theresulting solid particles are associated with a hydrophobic medium. Forexample, the solid particles may be suspended or immersed in ahydrophobic medium

In different embodiments of the compositions described herein the mediumchain fatty acid salt may be in the same particle or in a differentparticle than that of the API. It was found that bioavailability of acargo compound was lower if the medium chain fatty acid was in adifferent particle than the therapeutic agent i.e. there was improvedbioavailability if the medium chain fatty acid salt and the cargocompound were dried after solubilization together in the hydrophilicfraction. It is believed that if the medium chain fatty acid salt andthe cargo compound are dried after solubilization together in thehydrophilic fraction then they are in the same particle in the finalpowder.

Medium chain fatty acid salts include those having a carbon chain lengthof from about 6 to about 14 carbon atoms. Examples of fatty acid saltsare sodium hexanoate, sodium heptanoate, sodium octanoate (also termedsodium caprylate), sodium nonanoate, sodium decanoate, sodiumundecanoate, sodium dodecanoate, sodium tridecanoate, and sodiumtetradecanoate. In some embodiments, the medium chain fatty acid saltcontains a cation selected from the group consisting of potassium,lithium, ammonium and other monovalent cations e.g. the medium chainfatty acid salt is selected from lithium octanoate or potassiumoctanoate or arginine octanoate or other monovalent salts of the mediumchain fatty acids. The inventors found that raising the amount of mediumchain fatty acid salt increased the bioavailability of the resultingformulation. In particular, raising the amount of medium chain fattyacid salt, in particular sodium octanoate, above 10% to a range of about12% to 15% increased the bioavailability of the therapeutic agents inthe pharmaceutical compositions described herein.

In general, the amount of medium chain fatty acid salt in thecompositions described herein may be from 10% up to about 50% by weightof the bulk pharmaceutical composition. For example, the medium chainfatty acid salt may be present at an amount of about 10%-50%, preferablyabout 11%-40% most preferably about 11%-28% by weight for example atabout 12%-13%, 13%-14%, 14%-15%, 15%-16%, 16%-17%, 17%-18%, 18%-19%,19%-20%, 20%-21%, 21%-22%, 22%-23%, 23%-24%, 24%-25%, 25%-26%, 26%-27%,or 27%-28% by weight of the bulk pharmaceutical composition. In otherembodiments the medium chain fatty acid salt may be present at an amountof at least about 11%, at least about 12%, at least about 13%, at leastabout 14%, at least about 15% at least about 16%, at least about 17%, atleast about 18%, at least about 19%, at least about 20%, at least about21%, at least about 22%, at least about 23%, at least about 24%, atleast about 25%, at least about 26%, at least about 27% or at leastabout 28% by weight of the bulk pharmaceutical composition. In specificembodiments the medium chain fatty acid salt (sodium, potassium, lithiumor ammonium salt or a mixture thereof) is present at about 12%-21% byweight of the bulk pharmaceutical composition preferably 11%-18% orabout 11%-17% or 12%-16% or 12%-15% or 13%-16% or 13%-15% or 14%-16% or14%-15% or 15%-16% or most preferably 15% or 16%. In specificembodiments the medium chain fatty acid salt (having a carbon chainlength of from about 6 to about 14 carbon atoms particularly 8, 9 or 10carbon atoms) is present at about 12%-21% by weight of the bulkpharmaceutical composition preferably 11%-18% about 11%-17% or 12%-16%or 12%-15% or 13%-16% or 13%-15% or 14%-16% or 14%-15% or 15%-16% ormost preferably 15% or 16%. In specific embodiments the medium chainfatty acid salt (for example salts of octanoic acid, salts of subericacid, salts of geranic acid) is present at about 12%-21% by weight ofthe bulk pharmaceutical composition preferably 11%-18% about 11%-17% or12%-16% or 12%-15% or 13%-16% or 13%-15% or 14%-16% or 14%-15% or15%-16% or most preferably 15% or 16%. In certain embodiments the mediumchain fatty acid salt is present in the solid powder at an amount of 50%to 90%, preferably at an amount of 70% to 80%.

One embodiment of the invention comprises a composition comprising asuspension which consists essentially of an admixture of a hydrophobicmedium and a solid form wherein the solid form comprises atherapeutically effective amount of a therapeutic agent and at least onesalt of a medium chain fatty acid, and wherein the medium chain fattyacid salt is not a sodium salt. The salt may be the salt of anothercation e.g. lithium, potassium or ammonium; an ammonium salt ispreferred.

Matrix Forming Polymer:

In certain embodiments the composition of the invention comprises asuspension which comprises an admixture of a hydrophobic medium and asolid form wherein the solid form comprises a therapeutically effectiveamount of a therapeutic agent, at least one salt of a medium chain fattyacid and a matrix forming polymer, and wherein the matrix formingpolymer is present in the composition at an amount of 3% or more byweight. In certain embodiments the composition comprises a suspensionwhich consists essentially of an admixture of a hydrophobic medium and asolid form wherein the solid form comprises a therapeutically effectiveamount of a therapeutic agent, at least one salt of a medium chain fattyacid and a matrix forming polymer, and wherein the matrix formingpolymer is present in the composition at an amount of 3% or more byweight. In particular embodiments the matrix forming polymer is dextranor polyvinylpyrrolidone (PVP). In particular embodiments thepolyvinylpyrrolidone is present in the composition at an amount of about2% to about 20% by weight, preferably at an amount of about 3% to about18% by weight, more preferably at an amount of about 5% to about 15% byweight, most preferably at an amount of about 10% by weight. In certainparticular embodiments the polyvinylpyrrolidone is PVP-12 and/or has amolecular weight of about 3000. Other matrix forming polymers have asimilar effect in the compositions of the invention; such matrix formingpolymers include ionic polysaccharides (for example alginic acid andalginates) or neutral polysaccharides (for example dextran and HPMC),polyacrylic acid and poly methacrylic acid derivatives and highmolecular weight organic alcohols (for example polyvinyl alcohol).

Protease Inhibitors:

It is generally accepted in the art of delivery of proteins,polypeptides and peptides that protease inhibitors normally have to beadded to the formulation to prevent degradation of the API. However inthe formulations of the instant invention it is not necessary to addprotease inhibitors. The formulations of the invention appear to conferstability of the therapeutic agent to protease degradation within thetime-frame of activity i.e. the formulations of the invention areapparently environment inhibitory for enzyme activity. Additionally, theinventors performed an experiment wherein the protease inhibitoraprotinin was added to a formulation and this had no beneficial effecton activity. A similar experiment was performed where the proteaseinhibitor ε-aminocaproic acid was added to a formulation and this toohad no beneficial effect on activity. Therefore, in some embodiments, apharmaceutical composition described herein is substantially free of aprotease inhibitor.

Hydrophilic Fraction:

In embodiments of the invention, the above compounds, including thetherapeutic agent and the medium chain fatty acid salt are solubilizedin an aqueous medium and then dried to produce a powder. The dryingprocess may be achieved for example by lyophilization or granulation.The powder obtained is termed the “hydrophilic fraction”. In thehydrophilic fraction water is normally present at an amount of less than6%.

Lyophilization may be carried out as shown in the Examples herein and bymethods known in the art e.g. as described in Lyophilization:Introduction and Basic Principles, Thomas Jennings, published byInterpharm/CRC Press Ltd (1999, 2002) The lyophilizate may optionally bemilled (e.g. below 150 micron) or ground in a mortar. During industrialproduction the lyophilizate is preferably milled before mixing of thehydrophilic fraction and the hydrophobic medium in order to producebatch-to-batch reproducibility.

Granulation may be carried out as shown in the Examples herein and bymethods known in the art e.g. as described in Granulation, Salman et al,eds, Elsevier (2006) and in Handbook of Pharmaceutical GranulationTechnology, 2^(nd) edition, Dilip M. Parikh, ed., (2005

Various binders may be used in the granulation process such ascelluloses (including microcrystalline celluloses), lactoses (e.g.lactose monohydrate), dextroses, starch and mannitol and other bindersas described in the previous two references.

Hydrophobic Medium:

Oil:

As described above, in the compositions of the invention describedherein the therapeutic agent and the medium chain fatty acid salt are inintimate contact or association with a hydrophobic medium. For example,one or both may be coated, suspended, immersed or otherwise inassociation with a hydrophobic medium. Suitable hydrophobic mediums cancontain, for example, aliphatic, cyclic or aromatic molecules. Examplesof a suitable aliphatic hydrophobic medium include, but are not limitedto, mineral oil, fatty acid monoglycerides, diglycerides, triglycerides,ethers, esters, and combinations thereof. Examples of a suitable fattyacid are octanoic acid, decanoic acid and dodecanoic acid, also C7 andC9 fatty acids and di-acidic acids such as sebacic acid and subericacid, and derivatives thereof. Examples of triglycerides include, butare not limited to, long chain triglycerides, medium chaintriglycerides, and short chain triglycerides. For example, the longchain triglyceride can be castor oil or coconut oil or olive oil, andthe short chain triglyceride can be glyceryl tributyrate and the mediumchain triglyceride can be glyceryl tricaprylate. Monoglycerides areconsidered to be surfactants and are described below. Exemplary estersinclude ethyl isovalerate and butyl acetate. Examples of a suitablecyclic hydrophobic medium include, but are not limited to, terpenoids,cholesterol, cholesterol derivatives (e.g., cholesterol sulfate), andcholesterol esters of fatty acids. A non-limiting example of an aromatichydrophobic medium includes benzyl benzoate.

In some embodiments of the compositions described herein, it isdesirable that the hydrophobic medium include a plurality of hydrophobicmolecules. In some embodiments of the compositions described herein thehydrophobic medium also includes one or more surfactants (see below).

In some embodiments of the compositions described herein, thehydrophobic medium also includes one or more adhesive polymers such asmethylcellulose, ethylcellulose, hydroxypropylmethylcellulose (HPMC), orpoly(acrylate) derivative Carbopol®934P (C934P). Such adhesive polymersmay assist in the consolidation of the formulation and/or help itsadherence to mucosal surfaces.

Surface Active Agents (Surfactants):

The compositions of this invention described herein can further includea surface active agent. For example, the surface active agent can be acomponent of the hydrophobic medium as described above, and/or thesurface active agent can be a component of a solid form as describedabove, for example in the solid form or particle that includes thetherapeutic agent.

Suitable surface active agents include ionic and non-ionic surfactants.Examples of ionic surfactants are lecithin (phosphatidyl choline), bilesalts and detergents. Examples of non-ionic surfactants includemonoglycerides, cremophore, a polyethylene glycol fatty alcohol ether, asorbitan fatty acid ester, a polyoxyethylene sorbitan fatty acid ester,Solutol HS15, or a poloxamer or a combination thereof. Examples ofmonoglycerides are glyceryl monocaprylate (also termed glycerylmonooctanoate), glyceryl monodecanoate, glyceryl monolaurate, glycerylmonomyristate, glyceryl monostearate, glyceryl monopalmitate, andglyceryl monooleate. Examples of sorbitan fatty acid esters includesorbitan monolaurate, sorbitan monooleate, and sorbitan monopalmitate(Span 40), or a combination thereof. Examples of polyoxyethylenesorbitan fatty acid esters include polyoxyethylene sorbitan monooleate(Tween 80), polyoxyethylene sorbitan monostearate, polyoxyethylenesorbitan monopalmitate or a combination thereof. The commercialpreparations of monoglycerides that were used also contain variousamounts of diglycerides and triglycerides.

Compositions described herein including a surface active agent generallyinclude less than about 12% by weight of total surface active agent(e.g., less than about 10%, less than about 8%, less than about 6%, lessthan about 4%, less than about 2%, or less than about 1%). In particularembodiments of the invention the total sum of all the surfactants isabout 6%.

Methods of Making Pharmaceutical Compositions and the CompositionsProduced:

Also included in the invention are methods of producing the compositionsdescribed herein. Thus one embodiment of the invention is a process forproducing a pharmaceutical composition which comprises preparing awater-soluble composition comprising a therapeutically effective amountof at least one therapeutic agent and a medium chain fatty acid salt (asdescribed above), drying the water soluble composition to obtain a solidpowder, and suspending the solid powder in a hydrophobic medium, toproduce a suspension containing in solid form the therapeutic agent andthe medium chain fatty acid salt, thereby producing the pharmaceuticalcomposition, wherein the pharmaceutical composition contains 10% or moreby weight of medium chain fatty acid salt.

One embodiment is a process for producing a pharmaceutical compositionwhich comprises providing a solid powder of a therapeutically effectiveamount of at least one therapeutic agent and a solid powder comprising amedium chain fatty acid salt, and suspending the solid powders in ahydrophobic medium, to produce a suspension containing in solid form thetherapeutic agent and the medium chain fatty acid salt, therebyproducing the pharmaceutical composition, wherein the pharmaceuticalcomposition contains 10% or more by weight of medium chain fatty acidsalt.

In one embodiment of the processes and compositions described herein,the water-soluble composition is an aqueous solution. In certainembodiments the drying of the water-soluble composition is achieved bylyophilization or by granulation. In the granulation process a bindermay be added to the water soluble composition before drying. In certainembodiments the drying step removes sufficient water so that the watercontent in the pharmaceutical composition is lower than about 6% byweight, about 5% by weight, about 4% by weight, about 3% or about 2% orabout 1% by weight. In certain embodiments of the processes andcompositions described herein the drying step removes an amount of waterso that the water content in the solid powder is lower than 6% or 5% or4% or 3% or preferably lower than 2% by weight. The water content isnormally low and the water may be adsorbed to the solid phase duringlyophilization i.e. the water may be retained by intermolecular bonds.In certain embodiments the water soluble composition additionallycomprises a stabilizer for example methyl cellulose. In preferredembodiments of the of the processes and compositions described hereinthe hydrophobic medium is castor oil or glyceryl tricaprylate orglyceryl tributyrate or a combination thereof and may additionallycontain octanoic acid; in certain embodiments the hydrophobic mediumcomprises an aliphatic, olefinic, cyclic or aromatic compound, a mineraloil, a paraffin, a fatty acid such as octanoic acid, a monoglyceride, adiglyceride, a triglyceride, an ether or an ester, or a combinationthereof. In certain embodiments of the processes and compositionsdescribed herein the triglyceride is a long chain triglyceride, a mediumchain triglyceride preferably glyceryl tricaprylate or a short chaintriglyceride preferably glyceryl tributyrate, and the long chaintriglyceride is castor oil or coconut oil or a combination thereof. Incertain embodiments of the processes and compositions described hereinthe hydrophobic medium comprises castor oil or glyceryl tricaprylate orglyceryl tributyrate or a combination or mixture thereof, and mayadditionally comprise octanoic acid. In certain embodiments of theprocesses and compositions described herein the hydrophobic mediumcomprises glyceryl tricaprylate or a low molecular weight ester forexample ethyl isovalerate or butyl acetate. In certain embodiments ofthe processes and compositions described herein the main component byweight of the hydrophobic medium is castor oil and may additionallycomprise glyceryl tricaprylate. In certain embodiments of the processesand compositions described herein the main component by weight of thehydrophobic medium is glyceryl tricaprylate and may additionallycomprise castor oil.

A basic formulation is provided as an embodiment wherein the hydrophobicmedium consists essentially of castor oil, glyceryl monooleate andglyceryl tributyrate; in a further embodiment of the basic formulationthe hydrophilic fraction consists essentially of therapeutic agent,PVP-12 and sodium octanoate.

A particular formulation is provided as an embodiment wherein thehydrophobic medium consists essentially of glyceryl tricaprylate, castoroil, glyceryl monocaprylate, and Tween 80, and the hydrophilic fractionconsists essentially of therapeutic agent (e.g. octreotide), PVP-12 andsodium octanoate. Another particular formulation is provided as anembodiment wherein the hydrophobic medium comprises glyceryltricaprylate, castor oil, glyceryl monocaprylate, and Tween 80, and thehydrophilic fraction comprises therapeutic agent (e.g. octreotide),PVP-12 and sodium octanoate. In certain embodiments the hydrophobicmedium consists essentially of glyceryl tricaprylate and in certainembodiments additionally contains castor oil and/or glycerylmonocaprylate.

In certain embodiments the composition comprises a suspension whichconsists essentially of an admixture of a hydrophobic medium and a solidform wherein the solid form comprises a therapeutically effective amountof a therapeutic agent and at least one salt of a medium chain fattyacid, and wherein the medium chain fatty acid salt is present in thecomposition at an amount of 10% or more by weight. In certainembodiments the hydrophobic medium consists essentially of castor oil,glyceryl monooleate and glyceryl tributyrate; or the hydrophobic mediumconsists essentially of glyceryl tricaprylate and glycerylmonocaprylate; or the hydrophobic medium consists essentially of castoroil, glyceryl tricaprylate and glyceryl monocaprylate. In certainembodiments the hydrophobic medium comprises a triglyceride and amonoglyceride and in certain particular embodiments the monoglyceridehas the same fatty acid radical as the triglyceride. In certain of theseembodiments the triglyceride is glyceryl tricaprylate and themonoglyceride is glyceryl monocaprylate. In certain embodiments themedium chain fatty acid salt in the water-soluble composition has thesame fatty acid radical as the medium chain monoglyceride or as themedium chain triglyceride or a combination thereof. In certain of theseembodiments the medium chain fatty acid salt is sodium caprylate (sodiumoctanoate) and the monoglyceride is glyceryl monocaprylate and thetriglyceride is glyceryl tricaprylate.

Many of the compositions described herein comprise a suspension whichcomprises an admixture of a hydrophobic medium and a solid form whereinthe solid form comprises a therapeutically effective amount of atherapeutic agent and at least one salt of a medium chain fatty acid,and wherein the medium chain fatty acid salt is present in thecomposition at an amount of 10% or more by weight. The solid form may bea particle (e.g., consist essentially of particles, or consists ofparticles). The particle may be produced by lyophilization or bygranulation.

In a particular embodiment the formulation consists essentially of asuspension which comprises an admixture of a hydrophobic medium and asolid form wherein the solid form comprises a therapeutically effectiveamount of a therapeutic agent and about 10-20% preferably 15% mediumchain fatty acid salt preferably sodium octanoate, and about 5-10%preferably 10% PVP-12; and wherein the hydrophobic medium comprisesabout 20-80%, preferably 30-70% triglyceride preferably glyceryltricaprylate or glyceryl tributyrate or castor oil or a mixture thereof,about 3-10% surfactants, preferably about 6%, preferably glycerylmonocaprylate and Tween 80 and about 1% water; in particular embodimentsthe therapeutic agent is present at an amount of less than 33%, or lessthan 25%, or less than 10%, or less than 1% or less than 0.1%. The solidform may be a particle (e.g., consist essentially of particles, orconsists of particles). The particle may be produced by lyophilizationor by granulation. In a particular embodiment the solid form may be aparticle and may be produced by lyophilization or by granulation.

In a further embodiment the formulation consists essentially of asuspension which comprises an admixture of a hydrophobic medium and asolid form wherein the solid form comprises a therapeutically effectiveamount of a therapeutic agent and about 10-20% preferably 15% mediumchain fatty acid salt preferably sodium octanoate and about 5-10%preferably 10% PVP-12; and wherein the hydrophobic medium comprisesabout 20-80%, preferably 30-70% medium or short chain triglyceridepreferably glyceryl tricaprylate or glyceryl tributyrate, about 0-50%preferably 0-30% castor oil, about 3-10% surfactants, preferably about6%, preferably glyceryl monocaprylate and Tween 80, and about 1% water;in particular embodiments the therapeutic agent is present at an amountof less than 33%, or less than 25%, or less than 10%, or less than 1% orless than 0.1%.

In a particular embodiment the formulation consists essentially of asuspension which comprises an admixture of a hydrophobic medium and asolid form wherein the solid form comprises a therapeutically effectiveamount of a therapeutic agent and about 15% sodium octanoate and about10% PVP-12; and wherein the hydrophobic medium comprises about 41%glyceryl tricaprylate, about 27% castor oil, about 4% glycerylmonocaprylate, about 2% Tween 80, about 1% water and 1% or lesstherapeutic agent; when the therapeutic agent is octreotide it ispresent at about 0.058%.

In another particular embodiment the formulation consists essentially asuspension which comprises an admixture of a hydrophobic medium and asolid form wherein the solid form comprises a therapeutically effectiveamount of a therapeutic agent and about 15% sodium octanoate and about10% PVP-12; and wherein the hydrophobic medium comprises about 68%glyceryl tricaprylate, about 4% glyceryl monocaprylate, about 2% Tween80, about 15% sodium octanoate, about 10% PVP-12, about 1% water andless than 1% therapeutic agent; when the therapeutic agent is octreotideit is present at about 0.058%.

One embodiment is a composition comprising a suspension which comprisesan admixture of a hydrophobic medium and a solid form wherein the solidform comprises a therapeutically effective amount of octreotide and atleast one salt of a medium chain fatty acid; in a further embodiment themedium chain fatty acid salt is present in the composition at an amountof 10% or more by weight, preferably 15% by weight; in a furtherembodiment the solid form additionally comprises a matrix-formingpolymer. In a further embodiment the matrix forming polymer is dextranor polyvinylpyrrolidone (PVP). In a specific embodiment the matrixforming polymer is polyvinylpyrrolidone and the polyvinylpyrrolidone ispresent in the composition at an amount of about 2% to about 20% byweight, preferably about 10% by weight. In a specific embodiment thepolyvinylpyrrolidone is PVP-12 and/or the polyvinylpyrrolidone has amolecular weight of about 3000. In specific embodiments the hydrophobicmedium consists essentially of glyceryl tricaprylate and the solid formadditionally consists of PVP-12 and sodium octanoate. In more specificembodiments the hydrophobic medium additionally consists of castor oilor glyceryl monocaprylate or a combination thereof and a surfactant. Infurther specific embodiments the hydrophobic medium consists of glyceryltricaprylate, glyceryl monocaprylate, and polyoxyethylene sorbitanmonooleate (Tween 80). In a further embodiment the solid form consistsessentially of octreotide, PVP-12 and sodium octanoate. In a particularembodiment the composition contains about 41% of glyceryl tricaprylate,about 27% castor oil, about 4% glyceryl monocaprylate, about 2% Tween80, about 15% sodium octanoate, about 10% PVP-12, about 1% water andabout 0.058% octreotide. In another particular embodiment thecomposition contains about 68% of glyceryl tricaprylate, about 4%glyceryl monocaprylate, about 2% Tween 80, about 15% sodium octanoate,about 10% PVP-12, about 1% water and about 0.058% octreotide.

In all the above formulations, the percentages recited are weight/weightand the solid form may be a particle (e.g., consist essentially ofparticles, or consists of particles). The particles may be produced bylyophilization or by granulation.

Under normal storage conditions, the therapeutic agent within theformulations of the invention is stable over an extended period of time.The chemical and physical state of the formulation is stable. Onceadministered to the intestine the therapeutic agent is protected fromdamage by the GI environment since the formulations are oil-based andtherefore a separate local environment is created in the intestine wherethe therapeutic agent is contained in oil droplets, which confersstability in vivo.

In certain embodiments the process produces a composition which consistsessentially of a therapeutic agent and a medium chain fatty acid saltand a hydrophobic medium. In embodiments of the invention the solidpowder (solid form) consists essentially of a therapeutic agent and amedium chain fatty acid salt. Further embodiments of the invention arepharmaceutical compositions produced by the process describe herein. Incertain pharmaceutical compositions the therapeutic agent is a protein,a polypeptide, a peptide, a glycosaminoglycan, a polysaccharide, a smallmolecule or a polynucleotide and in particular embodiments thetherapeutic agent is insulin, growth hormone, parathyroid hormone,teriparatide, interferon-alfa (IFN-α), a low molecular weight heparin,leuprolide, fondaparinux, octreotide, exenatide, terlipressin,vancomycin or gentamicin. Particular embodiments of the inventioncomprise an oral dosage form comprising the pharmaceutical composition,in particular an oral dosage form which is enteric coated. Furtherembodiments of the invention comprise a capsule containing thecompositions of the invention, and in various embodiments the capsule isa hard gel or a soft gel capsule, and generally the capsule isenteric-coated. Other embodiments of the invention comprise a rectaldosage form comprising the pharmaceutical composition, in particular asuppository, or a buccal dosage form. A kit comprising instructions andthe dosage form is also envisaged.

The therapeutic agent or medium chain fatty acid salt, or anycombination of therapeutic agent and other components, such as proteinstabilizers, can be prepared in a solution of a mixture (e.g., formingan aqueous solution or mixture) which can be lyophilized together andthen suspended in a hydrophobic medium. Other components of thecomposition can also be optionally lyophilized or added duringreconstitution of the solid materials.

In some embodiments, the therapeutic agent is solubilized in a mixture,for example, including one or more additional components such as amedium chain fatty acid salt, a stabilizer and/or a surface activeagent, and the solvent is removed to provide a resulting solid powder(solid form), which is suspended in a hydrophobic medium. In someembodiments, the therapeutic agent and/or the medium chain fatty acidsalt may be formed into a granulated particle that is then associatedwith the hydrophobic medium (for example suspended in the hydrophobicmedium or coated with the hydrophobic medium). In general, thecompositions described herein are substantially free of “membranefluidizing agents” such as medium chain alcohols.

“Membrane fluidizing agents” are defined as medium chain alcohols whichhave a carbon chain length of from 4 to 15 carbon atoms (e.g., including5 to 15, 5 to 12, 6, 7, 8, 9, 10, or 11 carbon atoms). For example, amembrane fluidizing agent can be a linear (e.g., saturated orunsaturated), branched (e.g., saturated or unsaturated), cyclical (e.g.,saturated or unsaturated), or aromatic alcohol. Examples of suitablelinear alcohols include, but are not limited to, butanol, pentanol,hexanol, heptanol, octanol, nonanol, decanol, undecanol, dodecanol,tridecanol, tetradecanol, and pentadecanol. Examples of branchedalcohols include, but are not limited to, geraniol, farnesol, rhodinol,citronellol. An example of a cyclical alcohol includes, but is notlimited to, menthol, terpineol, myrtenol, perillyl and alcohol. Examplesof suitable aromatic alcohols include, but are not limited to, benzylalcohol, 4-hydroxycinnamic acid, thymol, styrene glycol, and phenoliccompounds. Examples of phenolic compounds include, but are not limitedto, phenol, m-cresol, and m-chlorocresol.

If desired, the pharmaceutical composition may also contain minoramounts of non-toxic auxiliary substances such pH buffering agents, andother substances such as for example, sodium acetate and triethanolamineoleate.

In at least one embodiment, a therapeutic agent, such as a protein, maybe chemically modified to enhance its half-life in circulation. Forexample, the therapeutic agent may undergo a process such as pegylation.

In some embodiments the process for producing a pharmaceuticalcomposition comprises preparing a water-soluble composition comprising atherapeutically effective amount of at least one therapeutic agent and amedium chain fatty acid salt, drying the water soluble composition toobtain a solid powder, and dissolving the solid powder in a solutionconsisting essentially of octanoic acid, thereby producing thepharmaceutical composition, which is a solution. In some embodiments,the solid form may be a particle (e.g., consist essentially ofparticles, or consists of particles). In some embodiments, the particlemay be produced by lyophilization or by granulation. In some embodimentsof this process the octanoic acid is present in the composition at alevel of about 60% to about 90% or at a level of about 70 to about 85%preferably about 78%. In some embodiments of this process the fatty acidsalt is sodium octanoate; in further embodiments of this process themedium chain fatty acid salt is present in the composition at an amountof about 11% to about 40% by weight or at an amount of about 11% toabout 28% by weight or at an amount of about 15% by weight. In someembodiments of this process the composition additionally comprises amatrix forming polymer and in particular embodiments of this process thematrix forming polymer is dextran or polyvinylpyrrolidone (PVP); infurther embodiments of this process the polyvinylpyrrolidone is presentin the composition at an amount of about 2% to about 20% by weight or atan amount of about 5% to about 15% by weight, preferably at an amount ofabout 10% by weight. In certain embodiments of this process thepolyvinylpyrrolidone is PVP-12 and/or has a molecular weight of about3000. The composition may in addition include surfactants as describedabove. The pharmaceutical products of these processes are furtherembodiments of the invention e.g. a composition containing octanoic acidat a level of about 60% to about 90% or at a level of about 70 to about85% preferably about 78%; fatty acid salt, preferably sodium octanoate,present in the composition at an amount of about 11% to about 40% byweight or at an amount of about 11% to about 28% by weight or at anamount of about 15% by weight; matrix forming polymer e.g.polyvinylpyrrolidone, preferably PVP-12, present in the composition atan amount of about 2% to about 20% by weight or preferably an amount ofabout 5% to about 15% by weight, preferably at an amount of about 10% byweight; and surfactants as described above. There also may be smallquantities of other hydrophobic constituents as described above.

Capsules:

Preferred pharmaceutical compositions are oral dosage forms orsuppositories. Exemplary dosage forms include gelatin or vegetariancapsules like starch hydroxylpropyl-methylcellulose (“HPMC”) capsules,enteric coated, containing the bulk drug product. Capsules which may beused to encapsulate the compositions of this invention are known in theart and are described for example in Pharmaceutical Capsules edited byPodczech and Jones, Pharmaceutical Press (2004) and in Hard gelatincapsules today—and tomorrow, 2nd edition, Steggeman ed published byCapsugel Library (2002).

Additional Formulations:

The compositions of the invention may be formulated using additionalmethods known in the art, for example as described in the followingpublications: Pharmaceutical Dosage Forms Vols 1-3 ed. Lieberman,Lachman and Schwartz, published by Marcel Dekker Inc, New York(1989);Water-insoluble Drug Formulation 2^(nd) edition, Liu, editor, publishedby CRC Press, Taylor and Francis Group (2008); Therapeutic Peptides andProteins: Formulation, Processing and Delivery Systems, 2^(nd) editionby Ajay K. Banga (author) published by CRC Press, Taylor and FrancisGroup (2006); Protein Formulation and Delivery, 2^(nd) edition, McNallyand Hasted eds, published by Informa Healthcare USA Inc(2008); andAdvanced Drug Formulation to Optimize Therapeutic Outcomes, Williams etal eds, published by Informa Healthcare USA (2008).

The compositions of the invention may be formulated usingmicroparticulate technology for example as described in MicroparticulateOral Drug Delivery, Gerbre-Selassie ed., published by Marcel Dekker Inc(1994) and in Dey et al, Multiparticulate Drug Delivery Systems forControlled Release, Tropical Journal of Pharmaceutical Research,September 2008; 7 (3): 1067-1075.

Methods of Treatment:

The compositions described herein exhibit effective, enteral delivery ofan unaltered biologically active substance (i.e. a therapeutic agent)and thus, have many uses. For example, the compositions described hereincan be used in the treatment of diabetes.

In particular, insulin to treat and prevent subjects (patients)suffering from Type II diabetes (prophylaxis of diabetes), and to treatpatients suffering from dysglycemia, pre-diabetes and metabolic syndromeand other conditions, may be administered in accordance with one or moreembodiments of the invention. Metabolic syndrome is a combination ofmedical disorders that increase the risk of developing cardiovasculardisease and diabetes. Metabolic syndrome is a composite of differentsymptoms: (1) fasting hyperglycemia (insulin resistance, type IIdiabetes, etc); (2) decreased HDL cholesterol; (3) elevatedtriglycerides; (4) high blood pressure; (5) central obesity; and (6)proinflammatory state.

One embodiment of the invention is a method of treatment or preventionof a subject suffering from the above conditions where the amount ofinsulin sufficient to treat the condition is a low dose of insulinformulated within the compositions of the invention. Low dose insulin isprovided by less than 300 or less than 200 Units per capsule e.g. 40-200Units per capsule.

Terlipressin (or other vasopressin analogs) to treat subjects (patients)suffering from hepato-renal syndrome (HRS), including HRS I and II,bleeding esophageal varices, portal hypertension and other conditionsmay be administered in accordance with one or more embodiments of theinvention. Such terlipressin formulations may also be used for primaryand secondary prophylaxis of variceal bleeding. A composition of theinvention comprises a suspension which comprises an admixture of ahydrophobic medium and a solid form wherein the solid form comprises atherapeutically effective amount of terlipressin (or other vasopressinanalogues) and at least one salt of a medium chain fatty acid.

Exenatide to improve glycemic control in subjects suffering from Type IIdiabetes and to treat other conditions such as obesity and for use inweight management may be administered in accordance with one or moreembodiments of the invention.

Interferon-alfa for the treatment of subjects suffering from chronichepatitis C and chronic hepatitis B and to treat other conditionsincluding cancer may be administered in accordance with one or moreembodiments of the invention.

Copaxone to treat subjects suffering from multiple sclerosis and totreat other conditions including inflammatory diseases may beadministered in accordance with one or more embodiments of theinvention.

Desmopressin to treat subjects suffering from primary nocturnalenuresis, central diabetes insipidus (DI) or bleeding disorders (VonWillebrand Disease and Hemopilia A) may be administered in accordancewith one or more embodiments of the invention. Oral desmopressinpreparations known in the art suffer from extremely low oralbioavailability.

Octreotide was first synthesized in 1979, and is an octapeptide thatmimics natural somatostatin pharmacologically, though it is a morepotent inhibitor of growth hormone, glucagon and insulin than thenatural hormone. Octreotide or other analogs of somatostatin may beadministered in accordance with one or more embodiments of the inventionfor use in treating or preventing a disease or disorder in a subjectsuffering from a disorder such as acromegaly, abnormal GI motility,flushing episodes associated with carcinoid syndrome, portalhypertension, an endocrine tumor (such as carcinoids, VIPoma),gastroparesis, diarrhea, pancreatic leak or a pancreatic pseudo-cyst.The diarrhea may result from radiotherapy or may occur for example insubjects with vasoactive intestinal peptide-secreting tumors (VIPomas).In addition, patients that undergo pancreatic surgery may suffer fromsecretion of extrinsic pancreas and are vulnerable to developingpancreatic leak or pseudo-cysts which may be treated by octreotideproducts of the invention. Some preferred embodiments are directed to amethod of treating a subject having a disorder such as acromegaly,abnormal GI motility, flushing episodes associated with carcinoidsyndrome, portal hypertension, an endocrine tumor (such as carcinoids,VIPoma), gastroparesis, diarrhea, pancreatic leak or a pancreaticpseudo-cyst, which comprises administering to the subject a compositionof the invention, wherein the therapeutic agent is octreotide, in anamount sufficient to treat the disorder. Octreotide formulations of theinvention may also be used for primary and secondary prophylaxis ofvariceal bleeding, which may be caused by portal hypertension; thevarices may be gastric or esophageal. Other uses of octreotideformulations of the invention are in treatment of shock of hypovolemic(e.g. hemorrhagic) or vasodilatory (e.g. septic) origin, hepatorenalsyndrome (HRS), cardiopulmonary resuscitation and anesthesia-inducedhypotension. Other analogs of somatostatin may be used in the methodsand compositions in which octreotide is used.

Vancomycin (molecular weight 1449 Da) is a glycopeptide antibiotic usedin the prophylaxis and treatment of infections caused by Gram-positivebacteria. The original indication for vancomycin was for the treatmentof methycilin-resistant Staphylococcus aureus (MRSA). Vancomycin neverbecame first line treatment for Staphylococcus aureus, one reason beingthat vancomycin must be given intravenously. The prior art preparationsof vancomycin need to be given intravenously for systemic therapy, sincevancomycin does not cross through the intestinal lining. It is a largehydrophilic molecule which partitions poorly across the gastrointestinalmucosa. The only indication for oral vancomycin therapy is in thetreatment of pseudomembranous colitis where it must be given orally toreach the site of infection in the colon. Vancomycin for use in treatingor preventing infection in a subject may be administered orally to thesubject in accordance with one or more embodiments of the invention.Some preferred embodiments of the invention are directed to a method oftreating or preventing an infection in a subject which comprisesadministering to the subject a composition of the invention, wherein thetherapeutic agent is vancomycin, in an amount sufficient to treat orprevent the infection.

Gentamicin (molecular weight=478) is an aminoglycoside antibiotic, usedto treat many types of bacterial infections, particularly those causedby gram-negative bacteria. When gentamicin is given orally in the priorart formulations, it is not systemically active. This is because it isnot absorbed to any appreciable extent from the small intestine.

In addition, compositions of the invention also can be used to treatconditions resulting from atherosclerosis and the formation of thrombiand emboli such as myocardial infarction and cerebrovascular accidents.Specifically, the compositions can be used to deliver heparin or lowmolecular weight heparin or fondaparinux across the mucosal epithelia.

The compositions of this invention can also be used to treathematological diseases and deficiency states such as anemia and hypoxiathat are amenable to administration of hematological growth factors. Thecompositions of the invention can be used to deliver vitamin B12 in asubject at high bioavailability wherein the mucosal epithelia of thesubject lacks sufficient intrinsic factor. G-CSF may also beadministered in accordance with various embodiments. Additionally, thecompositions of this invention can be used to treat osteoporosis, suchas through enteral administration of PTH, teriparatide or calcitoninonce or twice or more daily.

Human growth hormone (hGH) to treat growth hormone deficiency inparticular in children may be administered in accordance with one ormore embodiments. In some preferred embodiments, a composition describedherein comprising growth hormone can be administered to a subject totreat or prevent metabolic and lipid-related disorders, e.g., obesity,abdominal obesity, hyperlipidemia or hypercholestrolemia. For example acomposition of the invention comprising growth hormone can beadministered orally to a subject thereby treating obesity (e.g.,abdominal obesity). In some preferred embodiments, a compositiondescribed herein comprising growth hormone is administered to a subjectto treat or prevent HIV lipodistrophy (AIDS wasting) or to treatPrader-Willi syndrome, growth disturbance due to insufficient secretionof growth hormone (e.g. associated with gonadal dysgenesis or Turnersyndrome), growth disturbance in prepubertal children with chronic renalinsufficiency, and as replacement therapy in adults with pronouncedgrowth hormone deficiency. Compositions of the invention comprisinggrowth hormone can be administered orally to a subject to promote woundhealing and attenuate catabolic responses in severe burns, sepsis,multiple trauma, major operations, acute pancreatitis and intestinalfistula. Many other conditions besides GH deficiency cause poor growth,but growth benefits (height gains) are often poorer than when GHdeficiency is treated. Examples of other causes of shortness which maybe treated with compositions of the invention comprising growth hormoneare intrauterine growth retardation, and severe idiopathic shortstature. Other potential uses of compositions of the inventioncomprising growth hormone include treatment to reverse or preventeffects of aging in older adults, to aid muscle-building and astreatment for fibromyalgia.

Some preferred embodiments are directed to a method of treating adisorder such as obesity, HIV lipodistrophy, metabolic disorder, orgrowth deficiency in a subject which comprises administering to thesubject a composition of the invention wherein the therapeutic agent(the effector) is growth hormone, in an amount sufficient to treat thedisorder.

Some preferred embodiments are directed to a method of treating a bonedisorder in a subject which comprises administering to the subject acomposition of the invention, wherein the therapeutic agent isteriparatide or parathyroid hormone, in an amount sufficient to treatthe bone disorder.

Some preferred embodiments are directed to a method of treating orpreventing a blood coagulative disorder in a subject which comprisesadministering to the subject a composition of the invention wherein thetherapeutic agent is heparin or a heparin derivative or fondaparinux, inan amount sufficient to treat or prevent the blood coagulative disorder.

Leuprolide (GnRH agonist) formulated in an embodiment of the inventionmay be delivered for treatment of female infertility (e.g. once or twicedaily dosage), prostate cancer and Alzheimer's disease.

One embodiment of the invention relates to a method of treating asubject suffering from a disease or disorder which comprisesadministering to the subject a composition of the invention in an amountsufficient to treat the condition. Another embodiment of the inventionrelates to compositions of the invention for use in treating a diseaseor disorder in a subject. Another embodiment of the invention relates tothe use of a therapeutic agent in the manufacture of a medicament by theprocess of the invention for the treatment of a disorder.

The dosage regimen utilizing the compounds is selected in accordancewith a variety of factors including type, species, age, weight, sex andmedical condition of the patient; the severity of the condition to betreated; the route of administration; the renal and hepatic function ofthe patient; and the particular compound or salt thereof employed. Anordinarily skilled physician or veterinarian can readily determine andprescribe the effective amount of the drug required to prevent, counteror arrest the progress of the condition. Oral dosages of the presentinvention, when used for the indicated effects, may be provided in theform of capsules containing 0.001, 0.0025, 0.005, 0.01, 0.025, 0.05,0.1, 0.25, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0 or 100, 200, 300,400, 500, 600, 700, 800 or 1000 mg of therapeutic agent.

Compounds of the present invention may be administered in a single dailydose, or the total daily dosage may be administered in divided doses oftwo, three, four, five or six times daily. In some embodiments, thecomposition is administered at a daily dose of from about 0.01 to about5000 mg/day, e.g., administered once daily (e.g., in the morning orbefore bedtime) or twice or more daily (e.g. in the morning and beforebedtime).

A representative product of the invention is an API-based formulationorally administered as enteric coated-capsules: each capsule containsAPI co-lyophilized with PVP-12 and sodium octanoate, and suspended in ahydrophobic (lipophilic) medium containing: glyceryl tricaprylate,glyceryl monocaprylate, and Tween 80; in another representative productof the invention castor oil is additionally present. The compositionsdescribed herein can be administered to a subject i.e. a human or ananimal, in order to treat the subject with a pharmacologically ortherapeutically effective amount of a therapeutic agent describedherein. The animal may be a mammal e.g. a mouse, rat, pig horse, cow orsheep. As used herein the term “pharmacologically or therapeuticallyeffective amount” means that amount of a drug or pharmaceutical agent(the therapeutic agent) that will elicit the biological or medicalresponse of a tissue, system, animal or human that is being sought by aresearcher or clinician.

The formulations of the invention allow incorporation of the therapeuticagent into the formulation without any chemical modification of thetherapeutic agent. Additionally, as shown above, many differenttherapeutic agents have been successfully formulated within theformulations of the invention, including polypeptides, nucleotides,small molecules and even medium size proteins. Furthermore, theformulations of the invention allow for high flexibility in loading ofthe therapeutic agent. Loading capacity is dependent on the therapeuticagent. To date, loading capacity limits have not been reached; howeverloading of up to 1.5% wt/wt (polypeptides) and 6% wt/wt (smallmolecules) has been achieved and higher loading up to 33% is envisaged.Finally, the formulations of the invention protect the cargo compoundsfrom inactivation in the GI environment due to for example proteolyticdegradation and oxidation.

The function and advantages of these and other embodiments will be morefully understood from the following examples. These examples areintended to be illustrative in nature and are not to be considered aslimiting the scope of the systems and methods discussed herein.

EXAMPLES Example 1 Formulations

A. Composition of an Insulin Formulation

Table 1A presents an example of a composition in accordance with one ormore embodiments. More specifically, this composition is an insulinformulation. Insulin was obtained from Diosynth Biotechnology; sodiumoctanoate and NaOH from Merck; MgCl₂, MC400, Span40, lecithin and castoroil from Spectrum; PVP-12 from BASF; ethyl isovalerate from Merck/Sigma;glyceryl tributyrate from Acros/Penta; and glycerol monooleate fromAbitec Corp.

TABLE 1A Ingredient % w/w Hydrophilic Insulin 0.417 Fraction NaOH 0.029MgCl₂ 0.104 PVP-12 2.083 Sodium octanoate 3.125 Methyl cellulose 0.104Hydrophobic Castor oil 52.858 Medium Glyceryl tributyrate 28.466 Ethylisovalerate 8.195 Glycerol monooleate 1.779 Lecithin 1.893 Span-40 0.946

B. A Formulation for Leuprolide:

Table 1B presents an example of a composition for an API (ActivePharmaceutical Ingredient) in accordance with one or more embodiments.More specifically, this composition is a leuprolide formulation.

TABLE 1B Ingredient % w/w Hydrophilic Leuprolide 0.072 Fraction NaOH0.038 MgCl₂ 0.137 PVP-12 2.740 Sodium octanoate 12.002 Methyl cellulose0.137 Water 0.605 Hydrophobic Span-40 1.21 Medium Lecithin 2.43Ethyl-isovalerate 10.52 Glycerol monooleate 2.28 Glyceryl tributyrate23.74 Castor Oil 44.09

C. A Formulation with Decreased Amount of Hydrophobic Medium (50% ofHydrophobic Medium)

Table 1C presents an example of a composition for an API in accordancewith one or more embodiments. More specifically, this composition is aformulation for dextran (FD4). The FD4 is FITC-labeled dextran with a MWof 4.4 kDa (Sigma, FD4) and this is the dextran which was usedthroughout the Examples unless stated otherwise. This particularformulation contains coconut oil (Sigma) instead of GTB.

TABLE 1C Ingredient % w/w Hydrophilic Dextran 0.939 Fraction NaOH 0.001MgCl₂ 0.235 PVP-12 4.693 Sodium octanoate 20.662 Methyl cellulose 0.235Water 1.071 Hydrophobic Span-40 1.04 Medium Lecithin 2.08Ethyl-isovalerate 9.01 Glycerol-monooleate 1.95 Coconut oil 20.33 Castoroil 37.75The above formulations are used for a wide variety of therapeutic agentsand give good bioavailability to the cargo compound in the animal modelsdescribed below. Note that the net amount of therapeutic agent may varyas appropriate in any of the formulations and there may be minorvariations in the formulations; for example NaOH is not always used;coconut oil may be used instead of glyceryl tributyrate; MgCl₂ is notalways used (e.g. with hGH it is not used); all ingredients may besubstituted as described above in the specification.

Example 2 Schematic Representation of Insulin Formulation Production

FIG. 1 illustrates a method of producing a composition in accordancewith one or more embodiments. For example, this method may beimplemented to make the compositions presented above in Example 1.

Example 3 The Combination of Solid Particles Containing Sodium Octanoateand Hydrophobic Medium is Critical for Permeation Activity

FIG. 2 presents data relating to serum insulin levels after rectaladministration to rats. Rats were anesthetized and were administered 100μL of bulk drug formulation containing an insulin dose of 328 μg/rat (9IU/rat). Blood samples were collected at 0, 3, 6, 10, 15, 25, 30, 40, 60and 90 minutes post administration and serum was prepared fordetermination of human insulin by an immunoassay kit with no crossreactivity between rat and human insulin.

Data is presented as MEAN±SD, n=5. The left panel of FIG. 2 relates toadministration of human insulin with sodium octanoate (Na—C8) or solidhydrophilic fraction suspended in water (solid particles in water). Theright panel of FIG. 2 relates to administration of full insulinformulation (solid particles in hydrophobic medium). Table 2 belowpresents a summary of AUC values calculated from the concentration vs.time curves.

TABLE 2 Test compound AUC_((0-∞)) Na—C8 5753 ± 3569 Solid particles inwater 4083 ± 2569 Insulin in formulation 280933 ± 78692  (Solidparticles in hydrophobic medium) Data are MEAN ± SD

The average exposure (expressed by AUC values) to insulin after rectaladministration of insulin-SCD was about 50-fold higher than the exposureafter administration without a hydrophobic medium. Minimal exposure wasdetected in rats administered insulin with sodium octanoate alone or aspart of the solid particles of the hydrophilic fraction (as listed inExample 1) suspended in water. These data demonstrate synergy betweensolid sodium octanoate and a hydrophobic medium.

Example 4 Intestinal Absorption of Insulin after GI Administration ofInsulin to Rats

FIG. 3 presents data relating to serum insulin levels and blood glucoselevels after rectal administration of insulin solution and insulin informulation to rats. Rats were anesthetized and administered 100 μL oftest article (insulin in formulation or insulin in PBS) containing aninsulin dose of 328 μg/rat (9 IU/rat). Blood samples were collected at0, 3, 6, 10, 15, 25, 30, 40, 60 and 90 minutes post administration.Glucose level was immediately determined with a glucometer and serum wasprepared for determination of human insulin by an immunoassay kit withno cross reactivity between rat and human insulin.

Glucose levels are presented as the percentage form basal levelsmeasured before administration (time 0). The data of FIG. 3 is presentedas MEAN±SD, n=5.

Levels of insulin (left panel on FIG. 3) and glucose (right panel ofFIG. 3) after rectal administration of human insulin solubilized in PBS(insulin solution) or incorporated in the formulation are presented.Insulin levels rose rapidly in rat serum after rectal administration ofinsulin in formulation. Maximal levels were measured within 6 minutespost administration and a gradual drop detected until reaching basallevels at about 90 min post administration. This sharp and significantrise in insulin was accompanied by a significant drop in glucose levelsreaching an average of 20% of the initial levels already at 30 min postadministration. By contrast, rectal administration of insulin in PBScaused only a very slight glucose reduction, which is identical to thatobserved following treatment with the PBS control alone.

Example 5 Insulin Absorption after Rectal Administration of Insulin inFormulation to Rats

FIG. 4 presents data relating to changes in blood glucose and seruminsulin concentrations following SC (subcutaneous) administration ofinsulin solution (at 20 μg/rat) and rectal administration of insulin informulation (at 328 μg/rat). Blood samples were collected at 0, 3, 6,10, 15, 25, 30, 40, 60 and 90 minutes post rectal administration and at0, 15, 30, 45, 60, 90 min, 2, 3, and 4 hours post SC administration.Glucose was immediately determined with a glucometer and insulin by animmunoassay kit. Glucose levels are presented as the percentage formbasal levels measured before administration (time 0). The data of FIG. 4is presented as MEAN±SD, n=5.

The levels of insulin absorption from rat colon after insulin informulation administration were compared to the levels of insulinabsorbed after SC administration. Insulin exposure was calculated fromthe area under the serum concentration versus time curve (AUC) and theactivity calculated as the relative bioavailability (rBA) according tothe following equation:rBA=(rectal AUC _((0-∞)) /SC AUC _((0-∞)))*(SC dose/rectal dose)

Insulin penetration into the bloodstream occurs during a narrow windowof time, generally within about 10 minutes of rectal insulin informulation administration. The rise in serum insulin is paralleled by afall in blood glucose levels.

In order to derive information about insulin bioavailability whenformulated insulin is presented into the colon, AUC_((0-∞)) wasdetermined for rectal and SC administration and the rBA value of humaninsulin was 29.4±3.4% with coefficient of variance (CV)=11.4%.

Rectal administration of various insulin-containing formulations wascarried out on hundreds of animals. The assay was further developed andqualified as a bioassay to support platform development and batchrelease tests with a linear range of 10-200 μg/rat, repeatability of 39%and intermediate precision of 33%.

The insulin formulation described herein was tested in five differentstudies using a total of 25 rats. The rBA was 34.1±12.6% with CV of28.9%.

Example 6 Insulin Absorption after Intra-Jejunal Administration ofInsulin in Formulation to Rats

The absorption target site of the orally administered platform of theinvention is generally the small intestine. To test the activity ofinsulin formulation in rat intestine, two major obstacles wereaddressed: 1. Enteric-coated capsules for rats are not available andtherefore stomach bypass enabling direct intra-jejunal administration isneeded. 2. Insulin is extensively metabolized by the liver; in humans50-80% of endogenous insulin, secreted by pancreatic β-cells, issequestered by the liver and therefore can not be detected in thesystemic circulation. Insulin administered via the intestinal route (byway of insulin formulation) mimics the endogenous route of insulin asthe intestinal blood flow is drained into the portal vein which leadsdirectly to the liver. Therefore to determine insulin absorbance, bloodsamples must be drawn from the portal vein (portal circulation, prior tothe liver) as well as the jugular vein (systemic circulation, after theliver).

A specialized rat model in which three different cannulas are surgicallyimplanted in anesthetized rats was developed: 1. Jejunal cannula—stomachbypass, enables insulin formulation administration, 2. Portal veincannula—blood sampling prior to the liver, determine insulin that crossthe GI wall into the blood, and 3. Jugular vein cannula—to determine thesystemic levels of insulin. Using this model, the bioavailability ofinsulin in formulation (rBA) was determined

FIG. 5 presents data from a representative study relating to insulinlevels in the portal and systemic circulations after intra-jejunaladministration of insulin control and insulin formulation to rats. Rats(8 rats per group) were anesthetized and their jejunum exposed byabdominal surgery. The jejunum containing intestinal loop was placed ongauze and kept moist and fully intact throughout the entire study. Atemporary cannula was inserted into the jejunum and formulated insulinwas administered. Blood was collected from both portal and jugular veinsat the same time points, with approximately 4 time points per rat. TheMEAN±SD value of each time point was used to create a plasmaconcentration vs. time curve. AUC was determined and rBA was calculated.

Insulin levels in both the portal and systemic circulation rosedramatically after intra-jejunum administration of insulin informulation. This is in contrast to the minimal insulin absorbancedetected when insulin control was administered. The window of absorptionwas short and insulin levels peaked by 6 minutes. This profile issimilar to that seen after rectal administration of formulated insulin(see above). Higher insulin levels were detected in the portal comparedto the systemic circulation, with rBA of 10.1% compared to 5.6%,respectively.

Example 7 Additional Formulations Comprising Various Cargo Compounds

Table 3A details the components of a range of dextran formulations whichwere prepared as described in the following Examples. The sodium capratewas obtained from Fluka/Sigma, the olive oil from Fluka, the octanoicacid from Sigma and the mineral oil from Acros.

TABLE 3A Cargo Dextran Formulation A B C D E F G H Ingredient (% w/w) (%w/w) (% w/w) (% w/w) (% w/w) (% w/w) (% w/w) (% w/w) Hydrophilic Cargo0.545 0.939 0.565 0.546 0.565 0.565 0.565 0.551 fraction NaOH 0.0010.001 0.001 0.001 0.001 0.001 0.001 0.001 MgCl₂ 0.136 0.235 0.141 0.1560.141 0.141 0.141 0.138 PVP-12 2.726 4.693 2.823 3.117 2.823 2.823 2.8232.754 Sodium 12.001 20.662 — — 9.002 9.002 9.002 12.125 octanoate Sodium— — 9.002 — — — — — caprate MC 400 0.136 0.235 0.141 0.156 0.141 0.1410.141 0.138 Water 0.622 1.071 0.507 0.159 0.507 0.507 0.507 0.661Hydrophobic Span40 1.21 1.04 1.25 1.38 1.25 1.25 1.25 — medium Lecithin2.42 2.08 2.50 2.76 2.50 2.50 2.50 — Ethyliso- 10.46 9.01 10.83 11.9610.83 10.83 10.83 11.23 valerate Glyceryl 2.27 1.95 2.35 2.60 2.35 2.352.35 — monooleate Glyceryl 23.62 20.33 24.46 24.29 24.46 24.46 24.4625.35 tributyrate Coconut oil — — — — — — — — Castor oil 43.86 37.7545.42 45.07 45.42 — — 47.08 Octanoic acid — — — 7.80 — — — — Mineral oil— — — — — 45.42 — — Olive oil — — — — — — 45.42 —

Table 3B details the components of a range of teriparatide acetate andleuprolide formulations which were prepared as described in thefollowing Examples. Teriparatide was obtained from Novetide, andleuprolide was obtained from Bambio.

TABLE 3B Cargo Teriparatide Leuprolide Formulation I J K L Ingredient (%w/w) (% w/w) (% w/w) (% w/w) Hydrophilic Cargo 0.118 0.118 0.050 0.050fraction NaOH — — 0.040 0.04 MgCl₂ 0.137 0.137 0.142 0.15 PVP-12 2.7402.740 2.838 2.99 Sodium 12.001 12.001 9.012 — octanoate Sodium — — —4.48 caprate MC 400 0.137 0.137 0.142 0.15 Water 0.605 0.605 0.489 0.33Hydrophobic Span40 1.214 1.214 1.26 1.32 medium Lecithin 2.428 2.4282.52 2.65 Ethyl-iso- 10.515 10.515 10.89 11.46 valerate Glyceryl 2.2832.283 2.36 2.49 monooleate Glyceryl 23.740 — 24.59 25.87 tributyrateCoconut oil — 23.740 — — Castor oil 44.082 44.082 45.66 48.04

Table 3C details the components of hGH formulations which were preparedas described and the following Examples. The hGH was obtained from PLR,Israel (GHP-24).

TABLE 3C Cargo hGH Formulation O P Ingredient (% w/w) (% w/w)Hydrophilic Cargo 0.298 0.303 fraction NaOH — — MgCl₂ — — PVP- 12 2.8362.738 Sodium 9.006 12.007 octanoate Sodium — — caprate MC 400 0.1420.137 Water 0.492 0.607 Hydrophobic Span40 1.257 1.213 medium Lecithin2.514 2.427 Ethyl- 10.885 10.508 isovalerate Glyceryl 2.363 2.281monooleate Glyceryl 24.575 23.725 tributyrate Coconut oil — — Castor oil45.633 44.054

The production process for all these above formulations is essentiallyas described in FIG. 1 and in Example 11.

Example 8 Effect of Dose of Sodium Octanoate Incorporated in Formulationon Formulation Activity

The effect of increasing the amount of sodium octanoate (Na—C8) in theformulation on the activity of the formulation was tested usingformulations containing dextran (average MW=4.4 kDa, FITC labeled) ascargo compound and different doses of Na—C8 namely, formulation A inTable 3A (which contains 12% sodium octanoate by weight) and similardextran formulations containing different Na—C8 doses: 9%, 6% and 3%respectively.

To test the activity of these formulations in the jejunum ofnon-anesthetized rats, a rat model was established in which twodifferent cannulas are surgically implanted in male Sprague-Dowley rats

-   -   1—Jejunal cannula to bypass the stomach and enable direct        formulation administration to the jejunum.    -   2—Jugular vein cannula to determine the systematic levels of the        administered dextran following jejunal administration. Rats are        allowed to recover for 4 days before the study and are deprived        of food for 18 hours before the start of the study.

FIG. 6 presents data from a study which determines FITC-labeled dextran(4.4 kDa) bioavailability in non-anesthetized rats followingintra-jejunal administration of formulations containing differentamounts of Na—C8 or FITC-labeled dextran solubilized with the Na—C8 insaline solution (control).

The bioavailability of the different dextran formulations and thecontrol was evaluated by administrating the different formulationsdirectly to the jejunum of non-anesthetized rats and measuring plasmadextran levels at 3, 6, 10, 25, 60 and 90 minutes post administration.Levels of plasma dextran following administration of dextran informulation or in saline were compared to the levels of plasma dextranafter intravenous administration. Exposure values, AUC (0-90), weredetermined for jejunal and intravenous administration and the absolutebioavailability (aBA) was calculated according to the followingequation:aBA=(jejunal AUC(0-90))/(iv AUC(0-90))*(iv dose/jejunal dose).Data are presented as Mean±SD (n≧5 rats per group).

The results show that increasing the amount of Na—C8 incorporated in theformulation improves the bioavailability of the dextran in adose-responsive manner, reaching almost 30% aBA at the 12% (w/w) dose.Dextran administered with Na—C8 at similar doses and suspended in asaline solution (i.e. not formulated) showed much lower bioavailability(˜6% aBA). Further results dose-response results are shown in Example26.

Example 9 Effect of the Ratio of Hydrophilic Fraction/Hydrophobic Mediumon Formulation Activity

The effect on formulation activity of changing the ratio (weight/weight)between the hydrophilic fraction and the hydrophobic medium was testedusing formulations containing dextran (average MW=4.4 kDa, FITC labeled)as cargo (formulations A and B in Table 3A). The in vivonon-anesthetized rat model described in Example 8 was used in order tocompare the activity of the described formulations.

Table 4 presents bioavailability data following intra-jejunaladministration of formulations comprising a different ratio ofhydrophilic fraction to hydrophobic medium.

TABLE 4 Weight ratio between hydrophilic/ Animal Route of CargoFormulation hydrophobic medium model administration N % aBA ± SD DextranA 1/5.2 Rat Non- Jejunal 17 28.0 ± 6.8 B 1/2.6 anesthetized 19 24.8 ±25 

Formulations A and B were administered directly to the jejunum ofnon-anesthetized rats and plasma dextran levels were measured at 3, 6,10, 25, 60 and 90 minutes post formulation administration. The levels ofdextran absorption from rat jejunum after administration of dextran informulation were compared to the levels of dextran absorbed afterintravenous administration. Exposure values, AUC (0-90), were determinedfor jejunal and intravenous administration and the absolutebioavailability (aBA) determined according to the following equation:aBA=(jejunal AUC(0-90))/(iv AUC(0-90))*(iv dose/jejunal dose).

Data are presented as Mean±SD (n≧5 rats per group).

The results show that changing the ratio between the hydrophilicfraction and the hydrophobic medium in these formulations with a low %weight of therapeutic agent had no significant effect on thebioavailability of the cargo which gives a loading flexibility indevising additional formulations.

Example 10 Activity of Formulations Containing Different Cargo Compounds

In order to test the capability of the formulation platform, theactivity of formulations containing three different cargo compounds(APIs) was tested in three different animal models: jejunaladministration to non-anesthetized rats, rectal administration toanesthetized rats and jejunal administration to non-anesthetized pigs.Table 5 summarizes the results of representative experiments testing thebioavailability of formulations containing different APIs in the threedifferent animal models described above.

TABLE 5 Animal Route of API Formulation model Administration N % BA ±SDI Teriparatide I Rat non- Jejunal 5 14.0** ±10.8 anesthetized II I Pignon- Jejunal 5 15.0** ±9.3 anesthetized III Leuprolide K Rat non-Jejunal 4 10.1* ±7.5 anesthetized IV hGH P Rat Rectal 5 17.9** ±3.9anesthetized *Absolute BA (compared to IV) **Relative BA (compared toSC)

A. Leuprolide Absorption after Jejunal Administration of Leuprolide inFormulation to Rats

Table 5-III presents data from a representative study relating toleuprolide aBA following IV (intravenous) administration of leuprolidesolution (at 75 μg/Kg) and jejunal administration of leuprolide informulation (at 450 μg/Kg; formulation K, Table 3B) to non-anesthetizedrats, as previously described in Example 8.

Blood samples were drawn from the jugular vein at 3, 6, 10, 15, 25, 40,60 and 90 minutes post jejunal administration and at 3, 10, 25, 40, 90min, 2, 3.3 and 5 hours post IV administration, plasma was prepared andleuprolide levels were determined in each sample. Leuprolide levels insystemic circulation rose dramatically after jejunal administration ofleuprolide in formulation. Leuprolide blood levels peaked by 3 minutespost administration. The average aBA achieved after jejunaladministration of leuprolide in formulation was calculated as describedin the above Examples and was 10.1%. In a control experiment, jejunaladministration of leuprolide in PBS demonstrated negligible penetrationto the bloodstream.

A similar leuprolide formulation containing 12% sodium octanoate asdescribed in Table 1B was prepared; it was tested in the above model andshowed bioavailability as follows:rBA(compared to SC)=21.1%±12.0(CV=57%).

B. Teriparatide Absorption after Jejunal Administration of Teriparatidein Formulation to Rats

Table 5-I presents data from a representative study relating to plasmateriparatide concentration-time profiles following SC administration ofteriparatide solution (at 85 μg/formulation and jejunal administrationof teriparatide (teriparatide) in formulation (at 550 μg/Kg; formulationI, Table 3B) to non-anesthetized rats, as previously described inExample 8. Blood samples were drawn from the jugular vein at 3, 6, 10,25, 60 and 90 minutes post jejunal administration and at 3, 10, 30, 60,90 min, 2 and 3 hours post SC administration, plasma was prepared andteriparatide levels were determined in each sample. Teriparatide levelsin systemic circulation rose dramatically after jejunal administrationof teriparatide in formulation. Teriparatide levels peaked by 3 minutespost-administration. The average rBA achieved after jejunaladministration of teriparatide in formulation was calculated asdescribed in the above Examples, and was 14.0%. In a control experiment,jejunal administration of teriparatide in saline demonstrated nopenetration to the bloodstream.

C. Teriparatide Absorption after Jejunal Administration of Teriparatidein Formulation to Pigs

Table 5-II presents data from a representative study relating to plasmateriparatide concentration-time profiles following SC administration ofteriparatide solution (at 10.65 μg/Kg) and jejunal administration ofteriparatide in formulation (at 100 μg/Kg; formulation I, Table 3B) tonon-anesthetized pigs.

A pig model was established in which two different cannulas weresurgically permanently implanted in female domestic pigs:

1—jejunal cannula to bypass the stomach and enable direct formulationadministration to the jejunum.

2—jugular vein catheterization to determine the systematic levels of theadministered cargo following jejunal administration.

Pigs were allowed to recover for 7 days before the experiment anddeprived of food 18-20 hours before the start of the experiment.

Blood samples were drawn from the jugular vein at 0, 3, 6, 10, 15, 25,40, 60, 90 minutes, 2, 2.5 and 3 hours post jejunal administration andat 0, 3, 6, 10, 15, 20, 30, 45, 60, 90 min, 2, 2.5, 3 and 4 hours postSC administration, plasma was prepared and teriparatide levels weredetermined in each sample. Teriparatide levels in systemic circulationrose dramatically after jejunal administration of teriparatide informulation. Teriparatide levels peaked by 10 minutes postadministration. The average rBA achieved after jejunal administration ofteriparatide in formulation was calculated as described in the aboveExamples, and was 15.0%.

A similar pig experiment was performed using dextran (FD4, formulation Ain Table 3A) and it was determined that the average bioavailability ofdextran was 20% in pigs as compared to IV.

D. hGH Absorption after Rectal Administration of hGH in Formulation toRats

Table 5-IV presents data from a representative study relating to plasmahGH concentration-time profiles following SC administration of hGHsolution (at 81 μg/Kg) and rectal administration of hGH in formulation(at 800 μg/Kg; formulation P, Table 3C), to anesthetized rats.

Male Sprague-Dowley rats were deprived of food for 18 hours before thestart of the experiment. Rats were anesthetized by a solution ofketamine: xylazine. The formulation (100 μL/rat) was administeredrectally using a 14 G venflon. Blood samples were drawn from the jugularvein at 3, 6, 10, 15, 40, 60 and 90 minutes post rectal administrationand at 15, 30, 45, 60, 90 min, 2, 3, and 4 hours post SC administration,plasma was prepared and hGH levels were determined in each sample. hGHlevels in systemic circulation rose dramatically after rectaladministration of hGH in formulation. hGH levels peaked by 15 minutes.The average rBA achieved after rectal administration of hGH informulation was calculated as described in the above Examples and was17.9%. In a separate experiment hGH was administered to the jejunum andthe aBA was lower. In a control experiment, rectal administration of hGHin PBS demonstrated no penetration to the bloodstream.

Thus the results presented in Table 5 demonstrate that substantialexposure was obtained for all cargo compounds tested in all animalmodels tested.

The above results demonstrate that the formulations described hereinenable delivery of a wide range of different macromolecules through theintestinal epithelium in different animal models.

Example 11 Detailed Production Process of a Formulation of Teriparatide

Production of the Hydrophilic Fraction:

To 200 mL water the following ingredients were slowly added one by one(with 2-3 minutes mixing between each ingredient): 172 mg ofteriparatide, 200 mg of MgCl₂, 4.0 g of PVP-12, 17.52 g of sodiumoctanoate and 10.0 g of 2% MC-400 aqueous solution, prepared as follows:1 g of MC-400 powder was added to 50 mL water at 60±2° C. while mixing.After 5 min of mixing, the beaker was transferred to ice until a clearsolution was obtained.

After addition of the MC-400 solution, the solution was mixed foranother 5 min and then lyophilized for about 24 h. This procedureproduced about 22 g of hydrophilic fraction.

Production of the Hydrophobic Medium:

2 g of Span 40, 4 g of lecithin and 3.8 g of GMO were dissolved in 17.3g of ethyl isovalerate while mixing. To this solution were added 39.1 gof GTB and 72.6 g of castor oil. This procedure produced about 136-138 gof hydrophobic medium.

Production of the Bulk Drug Product:

Mixing of the hydrophilic fraction and the hydrophobic medium wasperformed at 20±2° C.

15.7 g of the hydrophilic fraction was slowly added during mixing to84.3 g of hydrophobic medium at 600±50 RPM. After addition of all thehydrophilic fraction, the mixing speed was increased to 2000±200 RPM for2-10 min followed by 4-8 cycles of 15 min mixing at 600±50 RPM and 2 minmixing at 2000±200 RPM.

Degassing by vacuum was then applied as follows: 5 min at 600 mBar, 5min at 500 mBar and 30-120 min at 400 mBar. The resulting suspension waspoured into a 100 mL dark bottle and stored at 2-8° C. This is theteriparatide formulation designated “I” described in Table 3B.

All other formulations described herein were produced by this method,varying ingredients and quantities according to the details given in therelevant Tables (see e.g. Example 29). A diagram of this method (withinsulin as cargo) is shown in FIG. 1.

Example 12 Effect of the Oil Incorporated in the Formulation onFormulation Activity

The effect of the type of oil incorporated in the formulation (in thehydrophobic medium) on formulation activity was tested. Formulationscontaining dextran (average MW=4.4 kDa, FITC labeled) as cargo compoundand different types of oils in the hydrophobic medium (formulations E, Fand G in Table 3A). were tested in rats.

To test the activity of these formulations in the jejunum ofnon-anesthetized rats, a rat model was established in which twodifferent cannulas are surgically implanted in male Sprague-Dowley rats:

-   -   1—Jejunal cannula to bypass the stomach and enable direct        formulation administration to the jejunum.    -   2—Jugular vein cannula to determine the systematic levels of the        administered dextran following jejunal administration.    -   Rats are allowed to recover for 4 days before the study and are        deprived of food for 18 hours before the start of the study.

Table 6 presents data from a study in non-anesthetized rats followingintra-jejunal administration of formulations containing different oilsin the hydrophobic medium.

TABLE 6 Cargo Formulation Oil N % aBA ± SD Dextran E Castor oil + GTB 1419.8 ± 5.5 F Mineral oil + GTB 5 12.2 ± 5.0 G Olive oil + GTB 5 12.0 ±9.9

Formulations containing different oils were administered directly to thejejunum of non-anesthetized rats and plasma dextran levels were measuredat 3, 6, 10, 25, 60 and 90 minutes post formulation administration. Thelevels of dextran absorption from rat jejunum after administration ofdextran in formulation were compared to the levels of dextran absorbedafter intravenous administration. Exposure values, AUC (0-90), weredetermined for jejunal and intravenous administration and the absoluteBioavailability (aBA) was determined according to the followingequation:aBA=(jejunal AUC(0-90))/(iv AUC(0-90))*(iv dose/jejunal dose).

Data are presented as Mean±SD (n≧5 rats per group).

Similar bioavailability was achieved when dextran was incorporated intoformulations containing castor oil or coconut oil. Good bioavailabilitywas also obtained in rat jejunum when teriparatide was used as cargocompound using formulations I and J; these formulations contain castoroil and GTB, and castor oil and coconut oil, respectively.

The results showed that formulations containing different kinds of oilsin their hydrophobic medium are active, enabling penetration of thecargo (dextran, teriparatide) carried by the formulation. Thus the datademonstrated that all tested oils enable bioavailability of the cargocarried by the formulation. Castor oil and coconut oil might be superiorto the other tested oils.

Example 13 Preparation of a Formulation Using Granulation Instead ofLyophilization

Production of the Hydrophilic Fraction:

To a plastic bag, the following ingredients were added: 1.00 g ofPVP-30, 6.70 g of sodium octanoate and 13.00 g of lactose monohydrate asbinder. After 5 min of mixing, all of the powder was transferred into amortar and pestle.

A dextran FD4 aqueous solution was prepared as followed: 0.42 g dextranwas dissolved in 1.2 g of WFI. All of the dextran solution was thenadded slowly to the powder while using a low shear agitation in a mortar& pestle; the agitation took around 45 min. The mixture was thentransferred into a lyophilization tray and was oven-dried for about 20 hat 50° C. This procedure produced about 20 g of hydrophilic fraction,which was a fine granulate.

Production of the Hydrophobic Medium:

2 g of Span 40, 4 g of lecithin and 3.8 g of GMO were dissolved in 17.3g of ethyl isovalerate while mixing. To this solution were added 39.1 gof GTB and 72.6 g of castor oil. This procedure produced about 136-138 gof hydrophobic medium.

Production of the Bulk Drug Product:

Mixing of the hydrophilic fraction and the hydrophobic medium wasperformed at 20±2° C.

19.00 g (29.58% of the final BDP) of the hydrophilic fraction was slowlyadded during mixing to 45.23 g (70.42% of the final BDP) of hydrophobicmedium at 600±50 RPM. After addition of all the hydrophilic fraction,the mixing speed was increased to 2000±200 RPM for 2-10 min followed by4-8 cycles of 15 min mixing at 600±50 RPM and 2 min mixing at 2000±200RPM.

Degassing by vacuum was then applied as follows: 5 min at 600 mBar, 5min at 500 mBar and 30-120 min at 400 mBar. The resulting suspension waspoured into a 100 mL dark bottle and stored at 2-8° C.

Rat Study:

The above suspension was administered rectally to rats as describedabove in the Examples and the results were as follows: 35% BA, 12.9% SD.Another batch of suspension prepared by granulation as described abovewas prepared and was administered to the jejunum of rats as describedabove in the Examples, and the results were as follows: 21.8% BA, 4.0%SD. A range of formulations are prepared in a similar manner usinggranulation and incorporating a selection of therapeutic agents andvarying the amount of sodium octanoate.

Example 14 Selection of Capsules

In vitro experiments were carried out using separately three types ofsolutions: the hydrophobic medium as described in the above Examples,ethyl isovalerate alone, and ethyl isovalerate containing 5% of each ofthe following surfactants: lecithin, span 40 and glyceryl mono-oleate. 3types of unsealed capsules, gelatin, starch and HPMC, were each filledwith each of these solutions. The filled capsules were then maintainedin vitro for 29 days at 22±2° C., 30-50% relative humidity. Gelatin andHPMC capsules gave the best results, namely no deformation of thecapsule.

Similar experiments were carried out using the same three solutions, andgelatin and HPMC capsules. The capsules were filled with the solutions,sealed (bonded) and then were maintained for 8 days at 22±2° C., 30-50%relative humidity. Both types of capsules showed stability to thesolutions tested i.e. there was no leakage and no deformation of thecapsules.

Example 15 Effect of Varying the Cation in the Medium Chain Fatty AcidSalt

Formulations were prepared with dextran (FD4) similar to Formulation Aof Table 3A except that 12% sodium octanoate (0.722M) was replaced by anequal molarity of lithium octanoate or potassium octanoate or arginineoctanoate (the last as a model for an ammonium salt). These formulationsare shown below in Table 7A.

TABLE 7A Formulation, cargo = dextran K-octanoate Li-octanoateArg-octanoate Ingredient (% w/w) (% w/w) (% w/w) Hydrophilic API 0.5450.546 0.546 fraction MgCl2 0.134 0.136 0.124 PVP-12 2.673 2.722 2.475Potassium 13.617 0.00 0.00 octanoate Lithium 0.00 10.826 0.00 octanoateArginine 0.00 0.00 22.989 octanoate MC 400 0.134 0.136 0.124 Water 0.6840.627 0.919 Hydrophobic Span40 1.185 1.206 1.097 medium Lecithin 2.3692.412 2.193 Ethyl 10.26 10.45 9.50 isovalerate Glyceryl 2.227 2.2682.062 monooleate Glyceryl 23.16 23.58 21.44 tributyrate Castor oil 43.0143.79 39.82

These formulations were each tested in the rat jejunal model describedin Example 8. The results were obtained and bioavailability wascalculated. The results are shown below in Table 7B.

TABLE 7B Medium chain fatty acid salt in formulation tested N % BA ± SDSodium octanoate (Formulation A) 18 22.2 ± 10.8 Lithium octanoate 11 8.4± 3.8 Potassium octanoate 10 7.9 ± 6.4 Arginine octanoate 12 17.5 ± 7.4 

The formulation A used in the above experiment was a different batch tothat used in Example 8, and so the BA results given here for formulationA differ slightly from those recited in Table 4.

The above results show that when 12% sodium octanoate was replaced inthe formulation by an equivalent molarity of lithium octanoate orpotassium octanoate, the formulation still had bioavailability but at alower level. The arginine octanoate formulation had similar activity tothe 12% sodium octanoate formulation.

Example 16 Effect of Addition of Medium Chain Alcohols (Geraniol andOctanol) to the Hydrophobic Medium

Formulations containing geraniol (BASF) and octanol (Spectrum/MP) wereprepared as described above, using the ingredients shown below in Table8. The sodium dodecanoate was obtained from Spectrum/Acros).

Formulation Q—low % Medium Chain Fatty Acid Salt:

A dextran (FD4) formulation was prepared essentially as described inExample 11, containing a total of 2.9% medium chain fatty acidsalt—(sodium octanoate 1.042%+sodium dodecanoate 1.869%)—and alsocontaining geraniol and octanol in the hydrophobic medium, all as shownin Table 8 below.

Formulation R—Over 10% Medium Chain Fatty Acid Salt:

A dextran formulation was prepared essentially as described forFormulation A except that geraniol and octanol were added to thehydrophobic medium, all as shown in Table 8.

TABLE 8 Formulation, cargo Dextran Dextran Q R Ingredient (% w/w) (%w/w) Hydrophilic API 0.545 0.456 fraction NaOH 0.029 0.000 MgCl₂ 0.1040.114 PVP-12 2.083 2.282 Sodium 1.042 10.046 octanoate Sodium 1.869 —dodecanoate MC 400 0.104 0.114 Water 0.231 0.521 Hydrophobic Geraniol9.148 8.39 medium Octanol 8.627 7.92 Span40 1.041 0.96 Lecithin 2.0811.91 Ethyl 9.012 8.27 isovalerate Glyceryl 1.956 1.80 monooleateGlyceryl 21.825 20.03 tributyrate Castor oil 40.532 37.20

Formulation Q (low % MCFA salt) was tested in the intra-jejunal ratmodel described above and the bioavailability was calculated: aBA=4.4%,SD=3.8 (n=12). Formulation R (over 10% MCFA salt) was tested in theintra-jejunal rat model described above and the bioavailability wascalculated: aBA=22.7%. SD=1.6 (n=6). The BA of these formulations do notdiffer significantly from similar formulations, described in the aboveExamples, which do not contain geraniol.

Example 17 Formulations for Gentamicin and for RNA

Formulations were prepared for gentamicin and for RNA essentially asdescribed in Example 11, with the ingredients of the bulk drug productas shown below in Table 9. The gentamicin was obtained from Applichemand the RNA was polyinosinic-polycytidylic acid sodium salt (Sigma).

TABLE 9A Formulation, API Gentamicin RNA Ingredient (% w/w) (% w/w)Hydrophilic API 6.000 0.100 fraction NaOH 0.670 — MgCl₂ 0.127 0.137PVP-12 2.545 2.741 Sodium octanoate 12.026 12.001 MC 400 0.127 0.137Water 0.860 0.605 Hydrophobic Span40 1.119 1.214 medium Lecithin 2.2382.429 Ethyl isovalerate 9.69 10.52 Glyceryl 2.103 2.283 monooleateGlyceryl 21.88 23.74 tributyrate Castor oil 40.62 44.09

The gentamicin formulation was tested in the rat jejunal model describedabove and in the rat rectal model described above (e.g. Examples 4 and5). The gentamicin was assayed using an immunoassay (ELISA). The resultsare shown in Table 9B below; % BA is calculated compared comparing to IVadministration. The formulations were shown to provide bioavailabilityto the gentamicin.

TABLE 9B % BA ± Cargo Formulation ROA N SD Gentamicin As Table 9Ajejunal 6 12.9 ± 4.5 As Table 9A rectal 5 50.1 ± 5.8

Similarly, the RNA formulation of Table 9A is tested in the rat jejunalmodel and in the rat rectal model described above. The RNA is assayedand the formulation is expected to provide bioavailability to the RNA.

Example 18 Effect on Formulation Activity of the Surfactants in theHydrophobic Medium

The effect on formulation activity of withdrawing surfactants from thehydrophobic medium was tested using formulations containing dextran(average MW=4.4 kDa, FITC labeled) as cargo (formulations A and H inTable 3A).

Table 10 presents data from a study in non-anesthetized rats followingintra-jejunal administration of formulations with or without surfactants(e.g. Span40, lecithin, glyceryl monooleate) in the hydrophobic medium.

TABLE 10 Surfactants in hydrophobic Cargo Formulation medium N % aBA ±SD Dextran A + 17 28.0 ± 6.8 H − 4 11.1 ± 8.2

Formulations with or without surfactants in the hydrophobic medium wereadministered directly to the jejunum of non-anesthetized rats and plasmadextran levels were measured at 3, 6, 10, 25, 60 and 90 minutes postformulation administration. The levels of dextran absorption from ratjejunum after administration of dextran in formulation were compared tothe levels of dextran absorbed after intravenous administration.

Exposure values, AUC (0-90), were determined for jejunal and intravenousadministration and the absolute bioavailability (aBA) was determinedaccording to the following equation: aBA=(jejunal AUC(0-90))/(ivAUC(0-90))*(iv dose/jejunal dose). Data are presented as Mean aBA±SD.

Lower bioavailability was achieved when dextran was incorporated into aformulation not containing surfactants in the hydrophobic medium(formulation H) as compared to a formulation containing surfactants inthe hydrophobic medium (formulation A). The results demonstrate thatwithdrawing surfactants from the hydrophobic medium adversely affectsformulation activity.

Example 19 Effect on Formulation Activity of Withdrawing Medium ChainFatty Acids from the Hydrophilic Fraction

The effect on formulation activity of withdrawing medium chain fattyacids (MCFA) from the hydrophilic fraction was tested using formulationscontaining dextran (average MW=4.4 kDa, FITC labeled) as cargo.

Table 11 presents data from a study in non-anesthetized rats followingintra-jejunal administration of formulations with or without sodiumoctanoate in the hydrophilic fraction (formulations A and D in Table 3A,respectively).

TABLE 11 MCFA in hydrophilic Cargo Formulation fraction N % aBA ± SDDextran A + 17 28.0 ± 6.8 D − 5  0.6 ± 1.0

The formulations described above were administered directly to thejejunum of non-anesthetized rats and plasma dextran levels were measuredat 3, 6, 10, 25, 60 and 90 minutes post formulation administration. Thelevels of dextran absorption from rat jejunum after administration ofdextran in formulation were compared to the levels of dextran absorbedafter intravenous administration. Exposure values, AUC (0-90), weredetermined for jejunal and intravenous administration and the absolutebioavailability (aBA) was determined according to the followingequation:aBA=(jejunal AUC(0-90))/(iv AUC(0-90))*(IV dose/jejunal dose).

Data are presented as Mean aBA±SD.

Negligible penetration of dextran was achieved when dextran wasincorporated into a formulation lacking medium chain fatty acids in thehydrophilic fraction (formulation D, % aBA=0.6±1.0) as compared to aformulation containing sodium octanoate at 12% w/w in the hydrophilicfraction (formulation A, % aBA=28.0±6.8). The results demonstrate that aformulation without medium chain fatty acids in the hydrophilic fractionis not active.

A similar experiment was performed using octreotide as cargo in theimproved formulation (see below). The rBA was 0.11% (CV=158%)

Example 20 Effect on Formulation Activity of Simplifying the Formulation

The effect on formulation activity of simplifying the formulation wastested using formulations containing dextran (average MW=4.4 kDa, FITClabeled) or octreotide (Novetide) as cargo. The basic formulationdescribed in the above Examples (e.g. formulations designated A, I andP) was simplified by not adding MgCl₂, and MC 400 to the hydrophilicfraction and by not adding span40, lecithin and ethyl iso-valerate tothe hydrophobic medium. There is a concomitant increase in the amountsof glyceryl monooleate (surfactant) and glyceryl tributyrate added tothe hydrophobic medium. Such formulations are shown in Table 12A below.These simplified formulations show no precipitation visually althoughthe particles are visible microscopically i.e. they are stablesuspensions.

TABLE 12A Formulation, API Dextran Octreotide Simplified SimplifiedIngredient (% w/w) (% w/w) Hydrophilic API 0.545 0.058 fraction NaOH0.001 0.000 MgCl₂ 0.000 0.000 PVP-12 2.735 2.750 Sodium octanoate 12.00012.019 MC 400 0.000 0.000 Water 0.611 0.593 Hydrophobic Span40 0.000.000 medium Lecithin 0.00 0.000 Ethyl isovalerate 0.00 0.000 Glycerylmonooleate 5.91 5.947 Glyceryl tributyrate 34.19 34.385 Castor oil 44.0044.248

The production process for these above simplified formulations isessentially as described in FIG. 1 and in Example 11 for the basicformulations.

The basic octreotide formulation is shown in Table 12B below.

TABLE 12B Cargo Octreotide Basic Formulation M Ingredient (% w/w)Hydrophilic Cargo 0.058 fraction NaOH 0.000 (HFP) MgCl₂ 0.137 PVP-122.742 Sodium 12.003 Octanoate MC 400 0.137 Water 0.603 HydrophobicSpan40 1.215 fraction Lecithin 2.430 (LFP) Ethyl-Iso- 10.522 valerateGlyceryl 2.284 Monooleate Glyceryl 23.756 Tributyrate Castor oil 44.113

Table 13 presents data from a study in non-anesthetized rats followingintra-jejunal administration of two different dextranformulations—formulation A of Table 3A and the simplified formulationshown in Table 12A.

TABLE 13 AUC (0-60 min)/ dose/kg Cargo Formulation N b.w. ± SD Dextran A(basic) 28 67062 ± 27368 Simplified 12 63897 ± 24210

The above results show that similar AUC values were achieved whendextran was incorporated into a formulation containing the basicformulation (formulation A) as compared to a simplified formulation.

Table 14 below presents data from a study in non-anesthetized ratsfollowing intra-jejunal administration of two different octreotideformulations—the basic formulation shown in Table 12B and the simplifiedformulation shown in Table 12A. The levels of octreotide absorption fromrat jejunum after administration of octreotide in basic formulation andsimplified formulation were obtained. Exposure values, AUC (0-25), weredetermined.

TABLE 14 AUC (0-25 min)/dose/kg Cargo Formulation N b.w. ± SD OctreotideBasic 13 2.8 ± 1.4 Simplified 13 2.3 ± 0.8

The above results in Table 14 show that the AUC values were slightlyless when octreotide was incorporated into a simplified formulation ascompared to the full formulation.

Example 21 Effect on Formulation Activity of Replacing Castor Oil byOctanoic Acid

The effect on formulation activity of replacing castor oil (and glyceryltributyrate and ethyl iso-valerate) by octanoic acid (Aldritch) wastested using a formulation containing dextran as cargo. This was done tomaintain the C8 motif in the formulation i.e. it was considered it mightbe advantageous to have C8 acid in the hydrophobic medium in addition tothe C8 salt in the hydrophilic fraction.

The effect of adding ricinoleic acid (Spectrum) was also tested bymaking a dextran formulation containing octanoic acid/ricinoleic acid.Ricinoleic acid was chosen since the main triglyceride component incastor oil is formed from ricinoleic acid. Three formulations of dextranwere prepared as shown in Table 15A below. The basic dextran formulationwas prepared essentially as described in the above Examples. The dextranoctanoic formulation was prepared essentially as described in the aboveExamples but wherein castor oil, glyceryl tributyrate and ethyliso-valerate were replaced by octanoic acid. This formulation was foundto be a solution by visual analysis but true solubility analysis was notperformed. It seems that the octanoic acid at high concentration (about78% of this formulation) dissolves the solid hydrophilic fraction, withthe PVP and sodium octanoate being soluble in octanoic acid at highconcentration. The dextran ricinoleic/octanoic acid formulation wasprepared essentially as described in the above Examples but whereincastor oil, glyceryl tributyrate and ethyl iso-valerate were replaced bya mixture of octanoic acid and ricinoleic acid. This formulation was asuspension as is usual for most of the formulations of this invention.

TABLE 15A Formulation, API Dextran Dextran Ricinoleic/ Dextran OctanoicOctanoic basic acid acid Ingredient (% w/w) (% w/w) (% w/w) HydrophilicAPI 0.545 0.545 0.545 fraction NaOH 0.001 0.001 0.001 MgCl₂ 0.136 0.1360.136 PVP-12 2.726 2.726 2.726 Sodium octanoate 12.001 12.002 12.002 MC400 0.136 0.136 0.136 Water 0.622 0.622 0.622 Hydrophobic Span40 1.2081.207 1.207 medium Lecithin 2.416 2.414 2.414 Ethyl isovalerate 10.460.00 0.00 Glyceryl monooleate 2.271 2.272 2.272 Glyceryl tributyrate23.62 0.00 0.00 Castor oil 43.86 0.00 0.00 Octanoic acid 0.000 77.9423.38 Ricinoleic acid 0.000 0.00 46.76 Ethyl Octanoate 0.000 0.00 7.80

The formulations described above in Table 15A were administered directlyto the jejunum of non-anesthetized rats, and plasma dextran levels weremeasured post formulation administration. Exposure values, AUC, weredetermined for the different formulations. These results are shown belowin Table 15B.

TABLE 15B AUC (0-60)/dose/kg Cargo Formulation N b.w. ± SD Dextran Basic12  72385 ± 37827 Octanoic acid 11 180824 ± 32778 Ricinoleic/Octanoicacid 11 113204 ± 33057

The results shown above in Table 15B demonstrate that the absorption ofdextran was much improved (over two-fold) in the formulation containingoctanoic acid. Additionally, the shape of the graph was changed showingslower but longer release. This may be advantageous since this allowsthe API to be longer-acting in the body. The dextran ricinoleic/octanoicresults showed less activity than the octanoic acid formulation, but wasstill improved over the basic formulation.

Since the octanoic acid and ricinoleic acid/octanoic acid formulationsshowed high activity, similar formulations were prepared with exenatideas cargo. Three formulations of exenatide were produced as shown inTable 16A below. The basic exenatide formulation was preparedessentially as described in the above Examples. The exenatide/octanoicformulation was prepared essentially as described in the above Examplesbut wherein castor oil, glyceryl tributyrate and ethyl iso-valerate werereplaced by octanoic acid. This formulation containing about 78%octanoic acid was found to be a solution by visual analysis, as was thesimilar dextran formulation above. The exenatide ricinoleic/octanoicacid formulation was prepared essentially as described in the aboveExamples but wherein castor oil, glyceryl tributyrate and ethyliso-valerate were replaced by a mixture of octanoic acid and ricinoleicacid.

TABLE 16A Formulation, API Exenatide Exenatide Ricinoleic/ ExenatideOctanoic Octanoic basic acid acid Ingredient (% w/w) (% w/w) (% w/w)Hydrophilic API 0.055 0.055 0.055 fraction NaOH 0.000 0.000 0.000 MgCl₂0.137 0.137 0.137 PVP-12 2.742 2.742 2.742 Sodium octanoate 12.00312.003 12.003 MC 400 0.137 0.137 0.137 Water 0.603 0.603 0.603Hydrophobic Span40 1.213 1.214 1.214 medium Lecithin 2.434 2.429 2.429Ethyl isovalerate 10.522 0.000 0.000 Glyceryl monooleate 2.283 2.2852.285 Glyceryl tributyrate 23.759 0.000 0.000 Castor oil 44.112 0.0000.000 Octanoic acid 0.000 78.395 47.035 Ricinoleic acid 0.000 0.00023.518 Ethyl Octanoate 0.000 0.000 7.842

The formulations described above in Table 16A were administered directlyto the jejunum of non-anesthetized rats, and plasma exenatide levelswere measured post formulation administration. Exposure values, AUC,were determined for the different formulations. These results are shownbelow in Table 16B.

TABLE 16B Cargo Formulation N AUC (0-90) ± SD % BA ± SD Exenatide Basic10 1961 ± 1791 8.8 ± 8.2 Octanoic acid 11 612 ± 350 3.1 ± 1.8 [AUC(0-180) ± SD] Ricinoleic/ 9 476 ± 321 2.2 ± 1.5 Octanoic acid

The results shown above in Table 16B demonstrate that the exenatideformulation containing octanoic acid showed bioavailability, but theabsorption of exenatide was decreased compared to the basic formulation.The shape of the graph was changed showing slower but longer release asin the case of the dextran octanoic acid formulation above; thisprolonged PK profile may be advantageous. Note that in the case of theoctanoic acid formulation, AUC 0-180 min was used for BA calculationsdue to the prolonged PK profile. The exenatide ricinoleic/octanoic acidformulation had even lower bioavailability than the octanoic acidformulation.

Example 22 Dose Response for Octanoic Acid

A. Octreotide Formulations:

The effect on formulation activity of varying the amount of octanoicacid was tested using formulations containing octreotide as cargo. Fourformulations of octreotide were prepared using 0%, 5%, 10% or 15%octanoic acid as shown in Table 17 below. The formulations are basicoctreotide formulations prepared essentially as described above whereinthe amount of octanoic acid varies as described and the amount of otheringredients in the hydrophobic medium. (ethyl isovalerate and glyceryltributyrate) was concomitantly reduced. (In these formulations thehydrophilic fraction was simplified to omit MgCl₂ and MC400.)

TABLE 17 Formulation, API Octreotide Octreotide Octreotide Octreotide 0%Octanoic 5% Octanoic 10% Octanoic 15% Octanoic Ingredient (% w/w) (%w/w) (% w/w) (% w/w) Hydrophilic API 0.058 0.057 0.057 0.057 fractionPVP-12 2.750 2.750 2.750 2.750 Sodium octanoate 12.019 12.034 12.03412.034 Water 0.593 0.594 0.594 0.594 Hydrophobic Span40 1.217 1.2191.219 1.219 medium Lecithin 2.441 2.437 2.437 2.437 Ethyl isovalerate10.554 0 0 0 Octanoic acid 0 5.053 10.553 15.021 Glyceryl monooleate2.290 2.291 2.291 2.291 Glyceryl tributyrate 23.832 29.325 23.825 19.357Castor oil 44.246 44.241 44.241 44.241

B. Exenatide Formulations:

The effect on formulation activity of varying the amount of octanoicacid was tested using formulations containing exenatide as cargo. Fiveformulations of exenatide were prepared using 0%, 10%, 15%, 20% or 35%octanoic acid as shown in Table 18 below. The formulations are basicexenatide formulations prepared essentially as described above whereinthe amount of octanoic acid varies as described and the amount of otheringredients in the hydrophobic medium (ethyl isovalerate and glyceryltributyrate) was concomitantly reduced.

TABLE 18 Formulation, API Exenatide Exenatide Exenatide ExenatideExenatide 0% 10% 15% 20% 35% Octanoic Octanoic Octanoic OctanoicOctanoic Ingredient (% w/w) (% w/w) (% w/w) (% w/w) (% w/w) HydrophilicAPI 0.055 0.055 0.055 0.055 0.055 fraction MgCl₂ 0.137 0.137 0.137 0.1370.137 PVP- 12 2.742 2.742 2.742 2.742 2.742 Sodium 12.003 12.003 12.00312.003 12.003 octanoate MC 400 0.137 0.137 0.137 0.137 0.137 Water 0.6030.603 0.603 0.603 0.603 Hydrophobic Span40 1.213 1.213 1.213 1.213 1.213medium Lecithin 2.434 2.434 2.434 2.434 2.434 Ethyl 10.522 0 0 0 0isovalerate Octanoic acid 0 10.522 15.081 20.085 34.282 Glyceryl 2.2832.283 2.283 2.283 2.283 monooleate Glyceryl 23.759 23.759 19.201 14.1970.000 tributyrate Castor oil 44.112 44.112 44.112 44.112 44.112

The formulations described above in Tables 17 and 18 above wereadministered directly to the jejunum of non-anesthetized rats, andplasma octreotide or exenatide levels were measured post formulationadministration. Exposure values, AUC, were determined for the differentformulations. These results are shown below in Table 19.

TABLE 19 AUC (0-60)/dose/kg Cargo Formulation N b.w. ± SD OctreotideBasic 14 2.8 ± 1.0 Basic 12 2.7 ± 1.2 5% Octanoic acid Basic) 12 3.2 ±1.2 10% Octanoic acid Basic 12 4.5 ± 2.3 15% Octanoic acid ExenatideBasic 10 3.9 ± 3.8 Basic, 10% Octanoic acid 15 4.6 ± 2.8 Basic, 15%Octanoic acid 6 3.0 ± 1.8 Basic, 20% Octanoic acid 5 2.2 ± 0.5 Basic,35% Octanoic acid 6 1.9 ± 0.7

The results shown above in Table 19 demonstrate that the octreotideformulation shows increased activity compared to the basic formulationas the amount of octanoic acid is increased to 15% (the maximum amounttested). Additionally, the results shown above in Table 19 demonstratethat the exenatide formulation shows increased activity compared to thebasic formulation as the amount of octanoic acid is increased to 15% andthe activity decreases at higher levels of octanoic acid.

Example 23 Effect of Different Medium Chain Fatty Acid Salts

A. Sodium Sebacate (Disodium Salt of Decanedioic Acid):

The effect on formulation activity of replacing sodium octanoate bysodium sebacate (disodium C10 salt) in a dextran formulation was tested.The sodium sebacate was prepared in situ from sebacic acid (Aldrich) andsodium hydroxide. The formulation produced is described in Table 20below. The formulation was prepared essentially as described above but12% sodium octanoate was replaced by sodium sebacate, at the same molarconcentration as sodium octanoate i.e. an equimolar amount of sodiumsebacate was used (viz., 0.72M).

TABLE 20 Formulation, API Dextran Na-Sebacate Ingredient (% w/w)Hydrophilic API 0.545 fraction NaOH 0.000 MgCl₂ 0.129 PVP-12 2.589Sodium Sebacate 16.190 MC 400 0.129 Water 0.783 Hydrophobic Span40 1.147medium Lecithin 2.295 Ethyl isovalerate 9.94 Glyceryl monooleate 2.157Glyceryl tributyrate 22.44 Castor oil 41.66

The formulation described above in Table 20 was administered directly tothe jejunum of non-anesthetized rats, and plasma dextran levels weremeasured post formulation administration. Exposure value, AUC, wasdetermined for the formulation and this is compared with a similarformulation prepared with sodium octanoate. These results are shownbelow in Table 21.

TABLE 21 AUC (0-60)/dose/kg Cargo Formulation N b.w. ± SD Dextran WithNa-octanoate 12 72385 ± 37827 With Na-Sebacate 9 18691 ± 11887

The results shown in Table 21 demonstrate that the dextran formulationcontaining sodium sebacate showed activity, but the absorption ofdextran was decreased compared to the formulation containing anequimolar amount of sodium octanoate.

B. Mono-Sodium Suberate or Di-Sodium Suberate

Octreotide-containing formulations were prepared wherein 12% sodiumoctanoate was replaced by an equimolar amount (0.72M) of mono-sodiumsuberate or of di-sodium suberate, which are C8 salts. These sodiumsalts were prepared in situ from suberic acid (Tokyo Chemical IndustryCo.) and sodium hydroxide.

TABLE 22A Formulation, API Octreotide Octreotide mono-Sodium di-Sodiumsuberate suberate (0.72M) (0.72M) Ingredient (% w/w) (% w/w) HydrophilicAPI 0.058 0.059 fraction PVP- 12 2.650 2.620 mono-Sodium 15.087 0Suberate di-Sodium 0 15.996 Suberate Water 0.712 0.747 HydrophobicSpan40 1.173 1.159 medium Lecithin 2.352 2.325 Ethyl 10.169 10.055isovalerate Glyceryl 2.206 2.181 monooleate Glyceryl 22.962 22.704tributyrate Castor oil 42.632 42.152

The formulations described above in Table 22 are administered directlyto the jejunum of non-anesthetized rats, and plasma octreotide levelsare measured post formulation administration. Exposure values, AUC, aredetermined for the formulations and this is compared with a similarformulation prepared with sodium octanoate.

C. Geranic Acid Salt

Two octreotide-containing formulations were prepared essentially asdescribed above wherein 12% sodium octanoate was replaced by 18% geranicacid sodium salt (0.95M) and 14.6% (0.77M) geranic acid sodium salt,which is 3,7-dimethyl-2,6-octadienoic acid (obtained from SAFC). Theformulations produced are described in Table 22B below.

TABLE 22B Formulation, API Octreotide Octreotide NaGeranate A NaGeranateB Ingredient (% w/w) (% w/w) Hydrophilic API 0.057 0.057 fraction NaOH 00.543 PVP 12 10.006 9.833 Sodium 18.053 14.625 Geranate Water 1.1831.084 Hydrophobic Tween 80 2.001 1.970 medium Glyceryl 4.001 3.923monocaprylate Glyceryl 63.235 65.927 tricaprylate Castor oil 0.000 0

The formulations described above in Table 22B were administered directlyto the jejunum of non-anesthetized rats, and plasma octreotide levelswere measured post formulation administration. Exposure values, AUC,were determined for the formulations and this was compared with asimilar formulation prepared with sodium octanoate. The results areshown below in Table 22C and they demonstrate that the formulation with18% sodium geranate had similar activity as the 12% sodium octanoateformulation, and the formulation with 14.6% sodium geranate hadincreased activity.

TABLE 22C AUC (0-60)/dose/kg Cargo Formulation N b.w. ± SD OctreotideSodium geranate A 9 4.48 ± 1.79 Sodium geranate B 9 6.33 ± 2.1  Improved9 4.38 ± 1.66

Example 24 Effect of PVP (Polyvinylpyrrolidone) on Formulation Activity

The effect on formulation activity of replacing PVP-12 by mannitol(Sigma) was tested using formulations containing exenatide as cargo. Itwas understood in the art that PVP-12 is a stabilizer and could bereplaced in the formulation by another stabilizer such as mannitol. Theformulation shown in Table 23 below was prepared. This formulation is abasic exenatide formulation prepared essentially as described above, butwherein PVP-12 is replaced by mannitol.

TABLE 23 Formulation, API Exenatide Mannitol Ingredient (% w/w)Hydrophilic API 0.055 fraction MgCl₂ 0.137 Mannitol 2.742 Sodium 12.003octanoate MC 400 0.137 Water 0.603 Hydrophobic Span40 1.213 mediumLecithin 2.434 Ethyl 10.522 isovalerate Glyceryl 2.283 monooleateGlyceryl 23.759 tributyrate Castor oil 44.112

The formulation described above in Table 23 was administered directly tothe jejunum of non-anesthetized rats, and plasma exenatide levels weremeasured post-formulation administration. Exposure values, AUC, weredetermined for the formulation compared to the basic formulation. Theseresults are shown below in Table 24.

TABLE 24 AUC (0-60)/dose/kg Cargo Formulation N b.w. ± SD ExenatideBasic 10 3.9 ± 3.8 Mannitol instead of PVP-12 6 1.6 ± 1.7

The results shown above in Table 24 demonstrate the surprising andunexpected result that the exenatide formulation without PVP-12 hadsignificantly decreased activity compared to the basic formulation. Itwas thus decided to investigate further the effect of PVP onbioavailability.

The effect on formulation activity of varying the molecular weight ofPVP was tested using formulations containing exenatide as cargo. Threeformulations of exenatide were prepared using either PVP-12, PVP-17 orPVP-25 (all obtained from BASF). PVP-12, PVP-17 and PVP-25 are allpolyvinylpyrrolidone polymers; the average molecular weights are about2500-3000, 10000 and 30000 respectively. The formulations are basicexenatide formulations prepared essentially as described above whereinthe PVP varies as described and wherein the hydrophilic fraction hasbeen simplified to omit MgCl₂ and MC400.

TABLE 25 Formulation, API Exenatide PVP- 12/17/25 Ingredient (% w/w)Hydrophilic API 0.022 fraction PVP 12/17/25 2.752 Sodium 12.005octanoate Water 0.602 Hydrophobic Span40 1.218 medium Lecithin 2.442Ethyl 10.561 isovalerate Glyceryl 2.291 monooleate Glyceryl 23.846tributyrate Castor oil 44.272

The three formulations described above in Table 25 were administereddirectly to the jejunum of non-anesthetized rats, and plasma exenatidelevels were measured post-formulation administration. Exposure values,AUC, were determined for the formulations. The results are shown belowin Table 26.

TABLE 26 AUC (0-60)/dose/kg Cargo Formulation N b.w. ± SD (a) PVP- 12 118.0 ± 7.7 Exenatide PVP- 17 6 3.4 ± 2.9 (b) PVP- 25 5 2.6 ± 2.3

The results shown above in Table 26 demonstrate that the exenatideformulations containing PVP-12 showed much higher activity than theexenatide formulations containing PVP-17 and PVP-25. Thus the effect ofPVP-12 only was investigated further, and it was decided to perform adose—response study using PVP-12. The effect of increasing the amount ofPVP-12 in the formulation on the activity of the formulation was testedusing formulations containing octreotide as cargo compound and differentdoses of PVP-12 as shown in Table 27 below. The PVP-12 doses tested were2.75% (the standard dose used in the above formulations) and 5.0%, 7.5%and 10.0% PVP-12; the hydrophilic fraction has been simplified to omitMgCl₂ and MC400. The formulation containing 10% PVP was semi-solid i.e.it was apparently a semi-solid suspension.

TABLE 27 Formulation, API Octreotide Octreotide Octreotide OctreotidePVP 2.75% PVP 5.0% PVP 7.5% PVP 10.0% Ingredient (% w/w) (% w/w) (% w/w)(% w/w) Hydrophilic API 0.058 0.057 0.057 0.057 fraction PVP- 12 2.7505.013 7.514 10.046 Sodium 12.019 12.031 12.037 12.018 octanoate Water0.593 0.684 0.784 0.885 Hydrophobic Span40 1.217 1.183 1.145 1.108medium Lecithin 2.441 2.373 2.297 2.222 Ethyl 10.554 10.259 9.934 9.608isovalerate Glyceryl 2.290 2.226 2.155 2.084 monooleate Glyceryl 23.83223.166 22.431 21.694 tributyrate Castor oil 44.246 43.009 41.645 40.278

The formulations described above in Table 27 were administered directlyto the jejunum of non-anesthetized rats, and plasma octreotide levelswere measured post-formulation administration. Exposure values, AUC,were determined for the four different formulations. These results areshown below in Table 28A.

TABLE 28A AUC (0-60)/dose/kg Cargo Formulation N b.w. ± SD Octreotide2.75% PVP- 12 14 2.8 ± 1.0 5.0% PVP- 12 12 3.7 ± 1.6 7.5% PVP- 12 12 4.2± 1.5 10.0% PVP- 12 11 4.7 ± 1.4

The results shown above in Table 28A demonstrate that the absorption ofoctreotide increased dramatically as the amount of PVP in theformulation increased. The formulation containing 10% PVP-12 hadabsorption of octreotide about 1.7 times greater that the formulationcontaining 2.75% PVP-12. An improved octreotide formulation in whichthere was 10% PVP-12 but no sodium octanoate showed virtually noactivity. The rBA was 0.11% (CV=158%) n=5.

It appears that the medium chain fatty acid salt acts as a permeabilityenhancer (by facilitating or enhancing permeability and/or absorption ofthe therapeutic agent), and that the PVP serves to increase the effectof the permeability enhancer in a synergistic manner since the PVP alonehas virtually no effect. See also Example 31.

A further experiment was performed to investigate if the 10% PVP-12could be replaced by dextran and still maintain activity of theformulation. The dextran was manufactured by Fluka; the averagemolecular weight is ˜6000. The formulations were prepared essentially asdescribed above wherein the PVP and dextran varies as described andwherein the hydrophilic fraction has been simplified to omit MgCl₂ andMC400 and where the sodium octanoate was increased to 15%; see Example26.

TABLE 28B Formulation, API Octreotide Octreotide Octreotide 10% Dextran5% Dextran 10% PVP no PVP no PVP Ingredient (% w/w) (% w/w) (% w/w)Hydrophilic API 0.058 0.058 0.058 fraction PVP- 12 10.011 0.0 0.0Dextran 0.0 10.011 5.011 Sodium 15.008 15.008 15.015 octanoate Water1.003 1.003 0.803 Hydrophobic Tween 80 2.027 2.027 2.169 medium Glyceryl4.036 4.036 4.319 monocaprylate Glyceryl 40.714 40.714 43.574tricaprylate Castor oil 27.143 27.143 29.049

The three formulations described above in Table 28B were administereddirectly to the jejunum of non-anesthetized rats, and plasma octreotidelevels were measured post-formulation administration. Exposure values,AUC, were determined for the formulations. The results are shown belowin Table 28C.

TABLE 28C AUC (0-25)/dose/kg Cargo Formulation N b.w. ± SD Octreotide10% PVP 9 4.4 ± 1.7 10% Dextran; no PVP 5 3.3 ± 1.6 5% Dextran; no PVP 93.2 ± 1.5

The results shown above in Table 28C demonstrate that the absorption ofoctreotide decreased when PVP in the formulation was replaced by dextranbut the activity was still significant. The formulation containing 10%dextran had absorption of octreotide about 75% of the formulationcontaining 10% PVP, and the formulation containing 5% dextran hadabsorption of octreotide about 73% of the formulation containing 10%PVP.

Example 25 A Comparative Study of C8, C9 and C10 Medium Chain Fatty AcidSalts viz., Sodium Octanoate, Sodium Nonanoate and Sodium Decanoate

The effect on formulation activity of replacing sodium octanoate withother medium chain fatty acid sodium salts was tested using formulationscontaining octreotide as cargo. Three formulations of octreotide wereprepared, as shown in Table 29 below. These are all basic formulationsprepared essentially as described above where the hydrophilic fractionhas been simplified to omit MgCl₂ and MC400 and wherein the medium chainfatty acid salt is an equimolar amount of sodium octanoate, sodiumnonanoate or sodium decanoate.

TABLE 29 Formulation, API Octreotide Octreotide Octreotide NaC8 12% NaC913% NaC10 14% (0.72M) (0.72M) (0.72M) Ingredient (% w/w) (% w/w) (% w/w)Hydrophilic API 0.058 0.057 0.058 fraction PVP- 12 2.750 2.718 2.685Sodium 12.019 0 0 octanoate Sodium 0 13.023 0 nonanoate Sodium 0 014.019 decanoate Water 0.593 0.632 0.670 Hydrophobic Span40 1.217 1.2031.188 medium Lecithin 2.441 2.412 2.383 Ethyl 10.554 10.428 10.303isovalerate Glyceryl 2.290 2.262 2.235 monooleate Glyceryl 23.832 23.54723.265 tributyrate Castor oil 44.246 43.718 43.194

The formulations described above in Table 29 were administered directlyto the jejunum of non-anesthetized rats, and plasma octreotide levelswere measured post-formulation administration. Exposure values, AUC,were determined for the formulations. The results are shown below inTable 30.

TABLE 30 AUC (0-25)/dose/kg Cargo Formulation N b.w. ± SD Octreotidesodium octanoate NaC8 9 2.1 ± 0.8 sodium nonanoate NaC9 10 2.5 ± 0.4sodium decanoate NaC10 10 1.7 ± 0.4

The results shown above in Table 30 demonstrate that when sodiumoctanoate in the formulation is replaced by sodium nonanoate or bysodium decanoate there is similar activity. Based on statisticalanalysis, there is no difference in activity between all threeformulations.

Example 26 Dose Response of Sodium Octanoate

The dose response of sodium octanoate at 12%, 15% and 18% was tested bymaking the formulations shown in Table 31. These are all basicformulations prepared essentially as described above where thehydrophilic fraction has been simplified to omit MgCl₂ and MC400 and thecargo compound was octreotide. Additionally the formulation wascorrected for viscosity i.e. the same or similar viscosity wasmaintained for all three formulations; this was achieved by varying theamounts of castor oil and glyceryl tributyrate.

TABLE 31 Formulation, API Octreotide Octreotide Octreotide NaC8 12% NaC815% NaC8 18% Ingredient (% w/w) (% w/w) (% w/w) Hydrophilic API 0.0580.058 0.058 fraction PVP- 12 2.750 2.652 2.554 (simplified) Sodium12.019 15.040 18.016 octanoate Water 0.593 0.710 0.825 HydrophobicSpan40 1.217 1.173 1.130 medium Lecithin 2.441 2.353 2.267 Ethyl 10.55410.175 9.802 isovalerate Glyceryl 2.290 2.207 2.126 monooleate Glyceryl23.832 32.816 41.090 tributyrate Castor oil 44.246 32.816 22.132

The formulations described above in Table 31 were administered directlyto the jejunum of non-anesthetized rats, and plasma octreotide levelswere measured post-formulation administration. Exposure values, AUC,were determined for the formulations. The results are shown below inTable 32.

TABLE 32 AUC (0-60)/dose/kg Cargo Formulation N b.w. ± SD OctreotideNaC8 12% 14 2.8 ± 1.0 NaC8 15% 12 4.1 ± 1.9 NaC8 18% 12 3.6 ± 1.1

The results shown above in Table 32 demonstrate that when sodiumoctanoate in the formulation is increased from 12% to 15% there is anincrease in activity but a further increase of sodium octanoate to 18%leads no higher activity than that obtained at 15%. Thus about 15%sodium octanoate appears to be the preferred amount.

Example 27 Investigation of the Effect of Varying theHydrophilic/Lipophilic Balance of the Surfactants in the Formulation

Table 33 below describes various octreotide formulations. The firstcolumn, formulation (a), is the basic formulation prepared essentiallyas described above where the hydrophilic fraction has been simplified toomit MgCl₂ and MC400, and the cargo compound is octreotide. Thesurfactants are Span 40, lecithin and glyceryl monooleate, and bycalculation the HLB is approximately 5-6. In the other formulations(formulations b, c and d) the HLB was changed as indicated (to 3.5, 6.7and 14) by replacing Span 40 and lecithin by differing amounts of Tween80 and by varying the amount of glyceryl monooleate.

TABLE 33 Formulation, API Octreotide Octreotide Octreotide OctreotideHLB HLB HLB HLB 5-6 [a] 3.5[b] 6.7[c] 14[d] Ingredient (% w/w) (% w/w)(% w/w) (% w/w) Hydrophilic API 0.058 0.057 0.057 0.057 fraction PVP-122.750 2.748 2.748 2.748 Sodium 12.019 12.027 12.027 12.027 octanoateWater 0.593 0.594 0.594 0.594 Hydrophobic Span40 1.217 0 0 0 mediumLecithin 2.441 0 0 0 Ethyl 10.554 10.547 10.546 10.547 isovalerate Tween80 0 0.502 2.003 5.500 Glyceryl 2.290 5.500 4.002 0.502 monooleateGlyceryl 23.832 23.811 23.811 23.811 tributyrate Castor oil 44.24644.215 44.215 44.215

The formulations described above in Table 33 were administered directlyto the jejunum of non-anesthetized rats, and plasma octreotide levelswere measured post-formulation administration. Exposure values, AUC,were determined for the formulations. The results are shown below inTable 34.

TABLE 34 AUC (0-25)/dose/kg Cargo Formulation N b.w. ± SD Octreotide HLB5-6 - [a] 9 2.1 ± 0.8 HLB 3.5 - [b] 12 3.3 ± 0.9 HLB 6.7 - [c] 11 3.8 ±0.9 HLB 14 - [d] 10 3.7 ± 0.9

The results shown above in Table 34 demonstrate that all the three newformulations replacing Span 40 and lecithin with Tween 80 [b, c and d]had much better activity than the basic formulation [a], even althoughthe HLB in [b] was lower, in [c] was slightly higher and in [d] was muchhigher than the HLB of the surfactants in (a). Additionally, theactivities of all the new formulations [(b, c, and d] were statisticallyvery similar. Thus the HLB alone of the surfactants does not seem toaffect activity but the characteristics of the surfactants appear toplay an important role. In particular, replacing Span 40 and lecithinwith Tween 80 is advantageous for activity in these octreotideformulations.

Example 28 Octreotide Formulations with Different Ratios of GlycerylTricaprylate to Castor Oil

Based on the accumulation of results described above including thePVP-12 dose response results, the sodium octanoate dose response resultsand the surfactant results inter alia, a series of octreotideformulations were prepared using 10% PVP-12 and 15% sodium octanoate,and varying the ratio of glyceryl tricaprylate to castor oil.Additionally, glyceryl monooleate and glyceryl tributyrate were replaced(if used) by glyceryl monocaprylate and glyceryl tricaprylate (bothsupplied by Abitec). This is to maintain the C8 motif within theformulation. Thus the hydrophilic fraction contains a salt of a C8 acid(octanoate) and the hydrophobic medium contains monoglycerides andtriglycerides incorporating the same C8 acid. The inventors believe thatthe use of C-8 compounds in both the hydrophilic fraction and in thehydrophobic medium may be advantageous for bioavailability. The amountsof Tween 80 and glyceryl monocaprylate were also varied in theformulations. The formulations were prepared are shown in Table 35Abelow. Formulations I, II, V and VI were semi-solid (apparentlysuspensions) and formulations III and IV were the usual liquidsuspensions.

TABLE 35A Formulation, API Octreotide Octreotide Octreotide OctreotideOctreotide Octreotide I II III IV V VI Ingredient (% w/w) (% w/w) (%w/w) (% w/w) (% w/w) (% w/w) Hydrophilic API 0.058 0.058 0.058 0.0580.058 0.058 fraction PVP- 12 10.011 10.011 10.011 10.011 10.011 10.011Sodium 15.008 15.008 15.008 15.008 15.008 15.008 octanoate Water 1.0031.003 1.003 1.003 1.003 1.003 Hydrophobic Tween 80 2.027 2.027 2.0272.027 6.063 6.062 medium Glyceryl 4.036 4.036 4.036 4.036 0 0monocaprylate Glyceryl 40.714 13.571 61.071 67.857 40.714 0 tricaprylateCastor oil 27.143 54.286 6.786 0.000 27.143 67.857

The formulations described above in Table 35A were administered directlyto the jejunum of non-anesthetized rats, and plasma octreotide levelswere measured post-formulation administration. Exposure values, AUC,were determined for the formulations. The results are shown below inTable 35B.

TABLE 35B AUC (0-25)/dose/kg Cargo Formulation N b.w. ± SD OctreotideFormulation I(GTC:castor oil 6:4) 9 4.4 ± 1.7 Formulation II(GTC:castoroil 2:8) 8 3.0 ± 1.7 Formulation III(GTC:castor oil 9:1) 9 3.1 ± 0.5Formulation IV(GTC:castor oil 10:0) 7 4.1 ± 2.1 Formulation V -withoutGMC 6 1.6 ± 1.0 (GTC:castor oil 6:4) Formulation VI -without GMC&GTC 71.1 ± 0.6 (GTC:castor oil 0:10)

The results shown above in Table 35B demonstrate that formulations 1 andIV have greatest activity. Since castor oil is absent in formulation IVthis demonstrates that castor oil is not essential for activity. Itseems that a high GTC:castor oil ratio e.g. 6:4 is beneficial foractivity. Additionally, since formulation V (which has low activity) hasthe same GTC:castor oil ratio as formulation I it appears thatadditionally GMC (or other monoglyceride) is desirable for activity.Additionally a formulation similar to formulation I of Table 36 wasprepared but sodium octanoate was omitted. This formulation showedvirtually no activity, rBA=0.1%.

Bulk drug product of formulation IV (improved, no castor oil) was milledwith a 150 micron screen, and then particle size was determined usingMalvern Laser Diffraction technology. Preliminary results indicated that90% (v/v) of the particles were below 130 microns, and 50% (v/v) of theparticles were below 45 microns.

Preliminary experiments using similar formulations to formulation I, butwith varying increased amounts of octreotide all gave similar BA i.e.there was approximately linear exposure independent of API loading. Apreliminary experiment using a similar formulation to formulation IV ateven higher octreotide loading—1.5% (wt/wt)—also gave similar BA.

A similar improved formulation to formulation I above was prepared usingFD4 as cargo instead of octreotide, and it was compared to a basicformulation. These formulations are described in Table 36A below.

TABLE 36A Formulation, API FD 4 Basic FD 4 (no Mg, MC) ImprovedIngredient (% w/w) (% w/w) Hydrophilic API 0.545 0.545 fraction NaOH0.001 0 PVP-12 2.734 10.012 Sodium 12.036 15.009 octanoate Water 0.6131.023 Hydrophobic Tween 80 0 2.013 medium Glyceryl 0 4.008 monocaprylateGlyceryl 0 40.434 tricaprylate Span40 1.21 0 Lecithin 2.42 0 Ethyl-Iso-10.49 0 valerate Glyceryl 2.28 0 mono-oleate Glyceryl 23.69 0tributyrate Castor oil 43.98 26.956

The formulations described above in Table 36A were administered directlyto the jejunum of non-anesthetized rats, and plasma FD4 levels weremeasured post-formulation administration. Exposure values, AUC, weredetermined for the formulations. The results are shown below in Table36B.

TABLE 36B AUC (0-90)/dose/kg Cargo Formulation N b.w. ± SD FD 4 Basic 667448 ± 16977 (dextran) Improved 6 95374 ± 47490

The results shown above in Table 36B demonstrate that the improvedformulation has much greater activity than the basic formulation.

Example 29 Detailed Production Process for a Selected (Improved)Octreotide Formulation

The octreotide formulation in Example 28 (Table 6, first column) wasprepared essentially as described in the above Examples. Below followsthe detailed production process for this formulation.

Production of the Hydrophilic Fraction:

To 150 mL water the following ingredients were slowly added and mixed:24.05 g of sodium octanoate, 16.04 g of PVP-12 and 92.4 g of 10 mg/mLaqueous octreotide solution. The resulting solution was lyophilized.

Production of the Hydrophobic Medium:

3.25 g Tween 80, 6.47 g of glyceryl monocaprylate, 65.25 g of glyceryltricaprylate and 43.50 g of castor oil were mixed together.

Production of the Bulk Drug Product:

26.08 g of the hydrophilic fraction was slowly added to 73.92 g of thehydrophobic medium at 20±2° C. while mixing. After addition of theentire hydrophilic fraction, the mixing speed was increased. Degassingby vacuum was then applied and the resulting suspension was stored at2-8° C.

To enable larger amounts of octreotide to be dissolved the followingmethod was devised:

-   -   1. The amount of water of the hydrophilic fraction preparation        was the same as the calculated volume of the final bulk drug        product.    -   2. PVP-12 was dissolved in half of the above amount of water.    -   3. Sodium octanoate was dissolved in the second half amount of        water.    -   4. Octreotide was dissolved in the PVP-12 solution (from        paragraph 2).    -   5. The sodium octanoate solution was added to the octreotide and        PVP-12 solution.

At this stage there was some precipitation, but it became soluble aftermixing.

Example 30 Experiments in Pigs Using Capsules

In order to test the activity of the formulations of the invention whenadministrated in capsules, an animal model allowing capsuleadministration to pigs (domestic swine) was established. In order tobypass the stomach and allow direct administration of capsules to thesmall intestine of the pig, a well established model in dogs (“NippleValve model”; Wilsson-Rahmberg & O. Jonsson, Laboratory Animals (1997),31, 231-240) was adapted to the commercial pig.

The two octreotide formulations shown below in Table 37 were prepared.The octreotide (x) formulation was prepared essentially as describedabove for the basic formulation wherein the hydrophilic fraction hasbeen simplified to omit MgCl₂ and MC400. The octreotide (y) formulationwas prepared essentially as described above for the improved octreotideformulation. The formulations were filled into gelatin capsules (fromCapsugel), basic formulation (x) at 0.42 mL/capsule and improvedformulation (y) at 0.44 mL/capsule, resulting in 5 mg net octreotidecontent in both types of filled capsules. The capsules were notenteric-coated i.e. they were uncoated.

TABLE 37 Formulation, API Octreotide (x) Octreotide(y) basic improvedIngredient (% w/w) (% w/w) Hydrophilic API 1.357 1.277 fraction PVP- 122.717 10.011 Sodium 12.011 15.008 octanoate Water 0.643 1.052Hydrophobic Tween 80 0 1.992 medium Glyceryl 0 3.967 monocaprylateGlyceryl 0 40.016 tricaprylate Castor oil 43.562 26.677 Span40 1.198 0Lecithin 2.403 0 Ethyl 10.391 0 isovalerate Glyceryl 2.254 0 monooleateGlyceryl 23.463 0 tributyrate

The formulations described above in Table 37 were administered directlyto the small intestine of the non-anesthetized pigs via the gastricbypass described above, and plasma octreotide levels were measuredpost-administration. Exposure values, AUC were determined for theformulations. The % BA was calculated compared to the exposure tooctreotide after subcutaneous administration. The results obtained areshown below in Table 38.

TABLE 38 Cargo Formulation N AUC (0-240) ± SD % BA ± SD OctreotideOctreotide(x) 4 896 ± 305 2.1 ± 0.7 Octreotide(y) 4 2574 ± 889  6.2 ±2.1

The above results in Table 38 show that there was bioavailability in thepig model for encapsulated formulations, for both the basic and improvedformulations. Octreotide bioavailability of the improved formulation wasabout three times the level of bioavailability of the basic formulation.

The results given here for bioavailability are underestimated becausesampling time was not sufficient for octreotide levels to go back tobaseline (0 ng/mL). This was due to the unexpectedly longer exposuretime in pigs as compared to what had been previously measured in rats.The shape of the graph was changed compared to the rat results showinglonger time to reach maximal peak levels and extended time in whichoctreotide is resident in the blood. This may be advantageous since thisallows the octreotide to be longer-acting in the body. Thus the actualbioavailability in pigs must be higher than the numbers given.

Based on the results in rats, the level of bioavailability in pigs ofoctreotide administered in aqueous solution is extrapolated to be about0.1%. This level of bioavailability is below the level of sensitivity ofthe bioassay used for pigs.

Example 31 Dose-Response Results for PVP in the Improved Formulation

Further to the PVP results in Example 24, the effect on activity ofincreasing the amount of PVP-12 in the improved formulation was studied.The improved formulations, made essentially as described above,contained octreotide as cargo compound and different doses of PVP-12 asshown in Table 39 below. The PVP-12 doses tested were 7.5%, 10.0% and15.0% PVP-12. The formulations containing 10% and 15.0% PVP weresemi-solid i.e. they were apparently semi-solid suspensions- and theformulation containing 7.5% PVP was a viscous suspension.

TABLE 39 Formulation, API Octreotide Octreotide Octreotide PVP 7.5% PVP10.0% PVP 15.0% Ingredient (% w/w) (% w/w) (% w/w) Hydrophilic API 0.0580.058 0.058 fraction PVP- 12 7.506 10.011 15.009 Sodium 15.012 15.00815.009 octanoate Water 0.903 1.003 1.203 Hydrophobic Tween 80 2.0982.027 1.884 medium Glyceryl 4.178 4.036 3.752 monocaprylate Glyceryl42.147 40.714 37.851 tricaprylate Castor oil 28.098 27.143 22.234

The formulations described above in Table 39 were administered directlyto the jejunum of non-anesthetized rats, and plasma octreotide levelswere measured post-formulation administration. Exposure values, AUC,were determined for the three formulations. These results are shownbelow in Table 40.

TABLE 40 T AUC (0-25)/dose/kg Cargo Formulation N b.w. ± SD Octreotide7.5% PVP-12 7 2.9 ± 2.2 10.0% PVP-12 9 4.4 ± 1.7 15% PVP-12 10 2.1 ± 1.2

The results shown above in Table 40 demonstrate that the absorption ofoctreotide was greatest when PVP in the formulation was 10%, andincreasing the amount to 15% results in significant decrease inactivity. This confirms the choice of 10% PVP in the improvedformulation.

Experiment 32 Activity of API Packed in Formulation Compared to APIAdministered Concomitant to Formulation

Three different basic formulations of three different cargo compoundswere prepared (dextran, gentamicin and exenatide), essentially asdescribed above (wherein the basic formulation is the basicnon-simplified hydrophilic fraction). Each of these three formulationswas administered directly to the jejunum of non-anesthetized rats, andplasma cargo levels were measured post-formulation administration.Exposure values, AUC, were determined for the formulations.Additionally, a similar formulation was prepared with a non-relevantcargo compound (a mock formulation). Separately, the mock formulationwas administered concomitantly with dextran, gentamicin or exenatide inaqueous solution and exposure values, AUC, were determined. Concomitantadministration was achieved by administrating cargo in aqueous solutionimmediately followed by mock formulation administration via ajejunal-implanted cannula (gastric bypass). For each compound, exposureafter administration of the formulated cargo was compared to exposureafter administration of the unformulated cargo (concomitant). Thecomparative results are shown below in Table 41. The results show thatthere is higher activity (bioavailability) when the cargo is formulatedcompared to unformulated (concomitant) in all three cases, and thatexenatide showed by far the greatest increase in activity due toformulation. Note that dextran and gentamicin are compounds that are notsensitive to protease degradation, whereas exenatide being a peptide issubject to degradation by intestinal enzymes. The large difference inactivity between the formulated exenatide compared to unformulatedexenatide may be due to the protective effect of the formulation againstdegradation.

TABLE 41 Formulated versus unformulated API/cargo (fold activity)Dextran 1.7 Gentamicin 1.5 Exenatide 4.4

Example 33 Intestinal Hyperpermeability Evaluation

A. Size Limitation:

The technology and formulations described above are intended to enhancethe permeability of the intestine, allowing specific delivery ofproteins, peptides and other otherwise impermeable molecules across thisbarrier. A certain degree of non-specific penetration of intestinalcontent may result as a side-effect of this enhancement of specificpermeability. The size of molecules which could possibly penetrate theintestine in a non-specific manner was evaluated using differentmolecular size markers.

In order to evaluate the molecular size limit of increased GIpermeability, five different FITC-labeled dextrans of differentmolecular weight were chosen to serve as molecular markers to testincreased intestinal permeability; the average molecular weight of thefive dextrans was 4.4, 10, 20, 40 and 70 kDa, equivalent to a radius of14, 23, 33, 45 and 60 Å respectively. These different size markers wereadministered directly to the jejunum of non-anaesthetized rats, throughan intestinal implanted cannula, and showed virtually no basalintestinal penetration when tested alone. Each of these markers was thenadministered directly to the jejunum of non-anesthetized rats togetherwith 300 μL of basic formulation, and the degree of its penetration wasevaluated by testing dextran levels in blood.

The plasma dextran levels were measured pre-dosing and at 3′, 6′, 10′,25′, 60′, 90′ minutes post formulation administration. Exposure values,AUC (0-90), were determined and the results are shown in FIG. 7. Data ispresented as MEAN±SD, n≧4.

The results show that while the smallest molecular marker tested(dextran of average MW=4.4 kDa), penetrates the intestine whenadministered concomitantly with a formulation, as the molecule sizeincreases, penetration extent decreases: a marker molecule of 10 kDapenetrates to a smaller extent and a 20 kDa marker to an even smallerextent. A marker molecule of 40 kDa shows minimal penetration, while amarker molecule of 70 kDa shows no penetration at all (basalpenetration). These results indicate that 40-70 kDa is a cutoff size fornon-specific permeability enhancement by formulations of the invention.Thus administration of a large volume of formulation (300 μL) to thejejunum of rats resulted in permeability enhancement of the intestinalbarrier, and this enhanced permeability is restricted by molecular size,showing a cutoff size of 40-70 kDa and minimal penetration at 40 kDa.

Published values of the size of hazardous molecules (molecular weightand radius) which could potentially be present in the intestine areshown below in Table 42.

TABLE 42 MW Radius (kDa) (Å) Macromolecules  >4 14 or larger LPS >100Short - 100 Long - 1000 Enterobacterial 70-900 — Toxins Viruses —600-1000 Bacteria — 10,000 or larger

Table 42 demonstrates that potentially hazardous molecules present inthe intestine are above the cutoff size of permeability enhancement bythe tested formulations, as shown above. Thus these results suggest thatthe tested formulations will not facilitate penetration of hazardousmolecules through the intestinal barrier and these formulations cantherefore be considered as safe. Other formulations of the inventiongive similar results.

B. Formulation Repeated Dosing:

In order to investigate if repeated dosing of formulation affectsintestinal permeability, the octreotide improved formulation (12% sodiumoctanoate with castor oil) was dosed to rats for 14 sequential daysusing the above in vivo model (rat implanted with two cannulas in thejejunum). At days 1, 7 and 14 of administration, a dextran permeabilitymarker (FITC-dextran of 4.4 kDa MW; FD4), was administered 60 minutespost formulation administration. This was to assess the permeability ofthe intestine by the penetration of the FD4 from the intestine to blood.No significant difference in FD4 exposure following 14 days offormulation repeated dosing was found. These results suggest there is noincrease in intestinal permeability following this period of repeateddosing of formulation, and intestinal enhanced permeability remains areversible process during this period.

The results suggest that the formulation causes no damage to theintestinal tissue, but acts by specifically opening the intestinalbarrier, showing no additive permeability enhancing effect.

Example 34 Intestinal Hyperpermeability Evaluation Time-Course andReversibility

Further to the study in the above Example, a study was designed in orderto define the time-course of increased intestinal permeability due tothe formulations of the invention, and the reversibility of thisprocess, using dextran as a permeability marker.

In order to define the time window of increased intestinal permeability,an in vivo model was developed in rats, in which one or two cannulas areimplanted in the jejunum of the rats. FITC-labeled dextran (averagemolecular weight 4.4 kDa, FD4), which has virtually no basal intestinalpenetration, served as a molecular marker to test intestinalpermeability. An experiment was designed in which the dextran marker wasadministered concomitant to the formulation (by a jejunal implantedcannula), or at different time intervals from the formulationadministration (by a second separate jejunal implanted cannula).Intestinal permeability was evaluated by testing FD4 penetration toblood. Rats were administered a basic formulation concomitant with thedextran marker, or the basic formulation and then the dextran marker atdifferent intervals of time (10, 30 and 60 minutes). Blood samples wereanalyzed for dextran concentration pre-administration and at 3, 6, 10,25, 60 and 90 min following dextran administration. The results areshown in FIG. 8. Data is presented as Mean±SD, n≧5.

FIG. 8 demonstrates that the dextran marker penetrates the intestine tothe highest extent when administered together with the formulation. Aninterval of 10 minutes between administration of the formulation andadministration of the dextran marker results in significantly decreasedamount of marker penetration, and increasing the interval furtherresults in exponential reduction of marker penetration.

These results show that while there is some degree of non-specificpermeability enhancing by the formulation, it is restricted to a shortperiod of time following administration of the formulation. Thepermeability of the intestine decreases sharply with time, and 60minutes from administration of the formulation there is no more markerpenetration. Thus administration of the formulation to the rat intestineresults in a very short period of hyperpermeability of the intestinalbarrier. Other formulations of the invention gave similar results.

Example 35 Oral Administration of Octreotide to Monkeys

In order to test the pharmacokinetics of octreotide following oraladministration of formulated octreotide to monkeys, five Cynomologusmonkeys were orally dosed with capsules containing an improved castoroil formulation of octreotide (similar to formulation I of Table 35—butwith higher load of octreotide). The capsules used were size 1 gelatincapsules coated with 6.7% Acryl-EZE® enteric coating; this coatingprevents capsule disintegration in the stomach and allows opening of thecapsules in the small intestine of the dosed animals. The octreotidedose used was 5 mg/capsule.

Monkeys were fasted overnight prior to capsule administration. Followingoral administration, blood samples were withdrawn over a period of 9.75hours, processed for plasma and analyzed for octreotide content by theLC/MS/MS method: see FIG. 9. Similar experiments were performed with theimproved no castor oil/GTC formulation (similar to formulation IV ofTable 35 but with higher load of API) and similar results were obtainedSimilar experiments were also performed with several different entericcoatings and similar results were obtained.

In order to compare the pharmacokinetics of octreotide followingadministration of the improved octreotide formulation, to thepharmacokinetics of injected octreotide, octreotide acetate solution(0.1 mg/monkey) was administered subcutaneously to two monkeys from theabove group to serve as a reference. Blood samples were withdrawn over aperiod of four hours, processed for plasma and analyzed for octreotidecontent by the LC/MS/MS method.

The pharmacokinetics of octreotide following oral octreotide andsubcutaneous injected octreotide solution were compared (see FIGS. 9 and10). The results of the oral formulation showed absorption over a periodof a few hours. The shape of the graph was changed compared tosubcutaneous, showing slower but longer release of octreotide into theblood. This may be advantageous since this allows the persistence ofoctreotide for a longer time in the blood potentially prolonging theactivity window.

An approved dose for injected octreotide acetate in humans is 0.1mg/patient. The above results in the monkeys suggest that the improvedformulation containing about 10 mg octreotide per dose will generatetherapeutic exposure in humans.

Example 36 Stability Data

Basic and improved octreotide formulations of the invention weremaintained both at 4° C. and at 25° C. and were tested for octreotidecontent periodically. Both formulations were found to be stable.

Example 37 Formulations Incorporating Vancomycin, Interferon-Alfa andTerlipressin

A. Vancomycin:

Table 43 below describes a vancomycin improved formulation, containing10% PVP and 15% sodium octanoate in the hydrophilic fraction, andcontaining glyceryl tricaprylate as the main constituent of thehydrophobic medium. The vancomycin was obtained from Gold Biotechnology.

TABLE 43 Formulation, API Vancomycin Ingredient (% w/w) Hydrophilic API6.267 fraction NaOH 0.082 PVP- 12 10.005 Sodium 15.016 octanoate Water1.216 Hydrophobic Tween 80 2.004 medium Glyceryl 4.008 monocaprylateGlyceryl 61.400 tricaprylate Castor oil 0.000

In a preliminary experiment, the formulation described above in Table 43was administered directly to the jejunum of non-anesthetized rats, andplasma vancomycin levels were measured post-formulation administration.Exposure value, AUC, was determined for the formulation. The results arethat the absolute BA is around 5% (comparative to IV, n=6). Whenvancomycin in saline solution was administered to the jejunum ofnon-anesthetized rats no BA was detected.

Interferon-Alfa:

Table 44 below describes an interferon-alfa improved formulation,containing 10% PVP and 15% sodium octanoate in the hydrophilic fraction,and containing glyceryl tricaprylate as the main constituent of thehydrophobic medium. The interferon-alfa is supplied in a buffer (fromIntas Biopharmaceuticals) and the ingredients of the interferon-alfabuffer in the formulation are marked by an asterisk (*).

TABLE 44 Formulation, API IFN-α Ingredient (% w/w) Hydrophilic API 0.050fraction *Na₂HPO₄ 0.032 *NaH₂PO₄ 0.030 *Polysorbate (Tween) 80 0.002*Disodium EDTA 0.002 PVP- 12 10.026 Sodium 14.997 Octanoate Water 1.006Hydrophobic Tween 80 2.005 medium Glyceryl 4.005 monocaprylate Glyceryl67.84 tricaprylate Castor oil 0

The formulation described above in Table 44 is administered directly tothe jejunum of non-anesthetized rats. Plasma interferon-alfa levels aremeasured post-formulation administration.

C. Terlipressin:

Table 45 below describes a terlipressin basic formulation and aterlipressin improved formulation containing 10% PVP and 15% sodiumoctanoate in the hydrophilic fraction, and containing glyceryltricaprylate as the main constituent of the hydrophobic medium. Theterlipressin was obtained from Bambio. The basic formulation wasprepared essentially as described above and the improved formulation isalso prepared essentially as described above.

TABLE 45 Formulation, API Terlipressin Terlipressin basic improvedIngredient (% w/w) (% w/w) Hydrophilic API 0.235 0.235 fraction MgCl₂0.137 0.000 PVP 12 2.736 10.004 Sodium 12.004 15.015 octanoate MC 4000.137 0.000 Water 0.610 1.010 Hydrophobic Span40 1.211 0.000 mediumLecithin 2.428 0.000 Ethyl 10.500 0.000 isovalerate Glyceryl 2.278 0.000monooleate Glyceryl 23.708 0.000 tributyrate Castor oil 44.016 0.000Tween 80 0.000 2.002 GMC 0.000 4.004 GTC 0.000 67.731

The formulations described above in Table 45 are administered directlyto the jejunum of non-anesthetized rats. Plasma terlipressin levels aremeasured post-formulation administration.

Example 38 Inhibition of Growth Hormone In Vivo by Octreotide

One of the best characterized effects of octreotide is the inhibition ofgrowth hormone release. In order to test for the efficacy of anoctreotide formulation of the invention on growth hormone inhibition, arat model was used in which endogenous rat growth hormone (rGH) levelswere monitored following octreotide formulation administration to thejejunum of the non-anesthetized rat model (described above).Administration of a basic octreotide formulation (containing 12% sodiumoctanoate) to the jejunum of rats was shown to reduce rGH levels by87.4% compared to administration of a saline control. This resultdemonstrates that the octreotide formulations described herein enabledelivery of octreotide in its active form from the intestinal lumen intothe blood stream.

Example 39 Toxicology Studies

A 28-day toxicity administration study of formulation control(excipients only, no cargo) was performed in Wistar rats. The animals inthe test group were daily administered rectally with the maximalfeasible dose of formulation (100 μL/animal/day) for 28 consecutivedays. The test group was compared to two control groups: a naïve group(non-treated) and a saline administered group, (n=15/group).

General clinical observations were made twice daily, and detailedclinical observations were performed weekly. Body weight and foodconsumption were measured weekly. Clinical pathology and gross pathologywere conducted one day after the last treatment. A histologicalexamination was performed on rectum, colon, liver and kidneys, and notoxic effects were detected. There was clean histopathology with nolocal GI or systemic findings, no formulation related clinical findings,no changes in hematological and blood chemistry parameters, nomacroscopic findings at necropsy and no mortality. In conclusion, thisexperiment demonstrated that there was no observed toxicity during adaily rectal dosing of formulation to rats for 28 consecutive days.

Having thus described several aspects of at least one embodiment, it isto be appreciated that various alterations, modifications, andimprovements will readily occur to those skilled in the art. Suchalterations, modifications, and improvements are intended to be part ofthis disclosure and are intended to be within the scope of theinvention. Accordingly, the foregoing description and drawings are byway of example only, and the scope of the invention should be determinedfrom proper construction of the appended claims, and their equivalents.

The invention claimed is:
 1. An enteric-coated oral dosage formcomprising a composition comprising a suspension which comprises anadmixture of a hydrophobic medium and a solid form wherein the solidform comprises a therapeutically effective amount of a polypeptide, atleast one salt of a medium chain fatty acid and polyvinylpyrrolidone(PVP), wherein the polyvinylpyrrolidone is present in the composition atan amount of 3% or more by weight, and wherein the at least one salt ofa medium chain fatty acid salt is present in the composition at anamount of at least 12% or more by weight.
 2. The enteric-coated oraldosage form of claim 1, wherein the medium chain fatty acid salt ispresent in the composition at an amount of 12% to 40% by weight.
 3. Theenteric-coated oral dosage form of claim 2 wherein the medium chainfatty acid salt is present in the composition at an amount of 12% to 18%by weight.
 4. The enteric-coated oral dosage form of claim 1 where thepolyvinylpyrrolidone is present in the composition at an amount of about3% to about 20% by weight.
 5. The enteric-coated oral dosage form ofclaim 4 where the polyvinylpyrrolidone is present in the composition atan amount of about 5% to about 15% by weight.
 6. The enteric-coated oraldosage form of claim 1, wherein the hydrophobic medium comprisesglyceryl tricaprylate and the solid form consists of a polypeptidepolyvinylpyrrolidone with a molecular weight of about 3000, and sodiumoctanoate.
 7. The enteric-coated oral dosage form of claim 1 wherein thehydrophobic medium additionally comprises castor oil or glycerylmonocaprylate or a combination thereof and a surfactant.
 8. Theenteric-coated oral dosage form of claim 1 wherein the hydrophobicmedium consists of glyceryl tricaprylate, glyceryl monocaprylate, andpolyoxyethylene sorbitan.
 9. The enteric-coated oral dosage form ofclaim 1 wherein the solid form consists essentially of polypeptide,polyvinylpyrrolidone with a molecular weight of about 3000, and sodiumoctanoate.
 10. The enteric-coated oral dosage form of claim 1 whereinthe composition comprises about 41% of glyceryl tricaprylate, about 27%castor oil, about 4% glyceryl monocaprylate, about 2% polyoxyethylenesorbitan monooleate, about 15% sodium octanoate, about 10%polyvinylpyrrolidone with a molecular weight of about 3000 and atherapeutically effective amount of polypeptide.
 11. The enteric-coatedoral dosage form of claim 1 wherein the composition comprises about 68%glyceryl tricaprylate, about 4% glyceryl monocaprylate, about 2%polyoxyethylene sorbitan monooleate, about 15% sodium octanoate, about10% polyvinylpyrrolidone with a molecular weight of about 3000 and atherapeutically effective amount of polypeptide.
 12. The enteric-coatedoral dosage form of claim 1, wherein the composition comprises atherapeutically effective amount of polypeptide, about 12-21% of sodiumoctanoate, about 5-10% of polyvinylpyrrolidone with a molecular weightof about 3000, about 20-80% of glyceryl tricaprylate, about 0-50% castoroil and about 3-10% surfactant.
 13. The enteric-coated oral dosage formof claim 1, wherein the composition comprises a therapeuticallyeffective amount of polypeptide, about 12-21% of sodium octanoate, about5-10% of polyvinylpyrrolidone with a molecular weight of about 3000,about 20-80% of glyceryl tricaprylate and about 3-10% surfactant. 14.The enteric-coated oral dosage form of claim 13, wherein the polypeptideis present at an amount of less than 33%.
 15. The enteric-coated oraldosage form of claim 14, wherein the composition comprises about 15% ofsodium octanoate, about 10% of polyvinylpyrrolidone with a molecularweight of about 3000, about 30-70% glyceryl tricaprylate and about 6% ofsurfactant.
 16. The enteric-coated oral dosage form of claim 15, whereinthe surfactant is glyceryl monocaprylate or polyoxyethylene sorbitanmonooleate.
 17. The enteric-coated oral dosage form of claim 1, whereinthe solid form comprises a particle or a plurality of particles.
 18. Theenteric-coated oral dosage form of claim 16, wherein the solid formfurther comprises a stabilizer.
 19. The enteric-coated oral dosage formof claim 1, wherein the solid form further comprises a stabilizer. 20.The enteric-coated oral dosage form of claim 13, wherein the polypeptideis present at an amount of less than 25%.
 21. The enteric-coated oraldosage form of claim 13, wherein the polypeptide is present at an amountof less than 10%.
 22. The enteric-coated oral dosage form of claim 13,wherein the polypeptide is present at an amount of less than 1%.
 23. Theenteric-coated oral dosage form of claim 13, wherein the polypeptide ispresent at an amount of less than 0.1%.
 24. The enteric-coated oraldosage form of claim 1, wherein the polyvinylpyrrolidone has a molecularweight of about
 3000. 25. The enteric-coated oral dosage form of claim1, wherein the medium chain fatty acid salt has a chain length fromabout 6 to about 14 carbon atoms.
 26. The enteric-coated oral dosageform of claim 25, wherein the medium chain fatty acid salt is sodiumhexanoate, sodium heptanoate, sodium octanoate, sodium nonanoate, sodiumdecanoate, sodium undecanoate, sodium dodecanoate, sodium tridecanoateor sodium tetradecanoate, or a corresponding potassium or lithium orammonium salt or a combination thereof.
 27. The enteric-coated oraldosage form of claim 26, wherein the medium chain fatty acid salt issodium octanoate.
 28. The enteric-coated oral dosage form of claim 1,wherein the hydrophobic medium comprises a mineral oil, a paraffin, afatty acid a monoglyceride, a diglyceride, a triglyceride, an ether oran ester, or a combination thereof.
 29. The enteric-coated oral dosageform of claim 1, wherein the hydrophobic medium comprises glyceryltricaprylate.
 30. The enteric-coated oral dosage form of claim 1,wherein the composition further comprises a surfactant.